Novel calcium channel drugs and uses

ABSTRACT

Novel multibinding compounds are disclosed. The compounds of this invention comprise 2-10 ligands covalently connected, each of the ligands being capable of binding to a ligand binding site in a Ca ++  channel , thereby modulating the biological activities thereof.

CONTINUING APPLICATION DATA

[0001] This application is a continuation-in-part of U.S. applicationSer. No. 09/325,557, filed Jun. 4, 1999, which is a continuation of U.S.application Ser. Nos. 60/088,465 (Jun. 8, 1998), Ser. No. 60/093,068(Jul. 16, 1998), and U.S. application Ser. No. 60,103,866 (Oct. 12,1998).

BACKGROUND

[0002] 1. Field of the Invention

[0003] This invention relates to novel multibinding compounds that bindto Ca⁺⁺ channels and modulate their activity. The compounds of thisinvention comprise 2-10 Ca⁺⁺ channel ligands covalently connected by alinker or linkers, wherein the ligands in their monovalent (i.e.unlinked) state bind to and are capable of modulating the activity ofone or more types of Ca⁺⁺ channel. The manner of linking the ligandstogether is such that the multibinding agents thus formed demonstrate anincreased biologic and/or therapeutic effect as compared to the samenumber of unlinked ligands made available for binding to the Ca⁺⁺channel. The invention also relates to methods of using such compoundsand to methods of preparing them.

[0004] The compounds of this invention are particularly useful fortreating diseases and conditions of mammals that are mediated by Ca⁺⁺channels. Accordingly, this invention also relates to pharmaceuticalcompositions comprising a pharmaceutically acceptable excipient and aneffective amount of a compound of this invention.

[0005] 2. State of the Art

[0006] Voltage-gated Ca⁺⁺ channels mediate the influx of Ca⁺⁺ into cellsin response to changes in membrane potential. Because of their centralroles in ion homeostasis and in cell signaling events, these channelsare involved in a wide variety of physiological activities, e.g., musclecontraction, cardiovascular function, hormone and neurotransmittersecretion, and tissue growth and remodeling processes.

[0007] At least six types of calcium channels have been identified andcharacterized (Table 1, Appendix). The high-voltage activated Ca⁺⁺channels are formed by the heteromeric association of membrane proteinscomprising at least three subunits α (α₁, α₂), δ, β (and γ in skeletalmuscle). The α₁ subunit alone is sufficient to form a functionalchannel, although the functional properties of the channel are subjectto modification, particularly by the β subunit. The α₁ subunit isorganized into four homologous domains (I-IV), each domain including 6transmembrane segments (S1-S6) (FIG. 1, Appendix). It is thought thatthe channel pore is formed from S5, S6 and the region between them, andthat the voltage sensor resides in S4.

[0008] These channels exist in resting (closed), activated (open ) orinactivated (desensitized) states. The resting channels open in responseto depolarization of the membrane, then transition to an inactivatedstate. Repolarization is required for return to the resting state. Asshown in Table 1, channels differ in their activation and inactivationproperties.

[0009] Not surprisingly, Ca⁺⁺ channels are recognized as importanttargets for drug therapy. They are implicated in a variety of pathologicconditions, including, e.g., essential hypertension, angina, congestiveheart failure, arrythmias, migraine and pain.

[0010] Calcium channel antagonists are potent vasodilators and arewidely used in the treatment of hypertension and angina pectoris. Thecompounds approved for clinical use in the U.S. fall into severalchemical classes: the dihydropyridines (e.g., amlodipine, felodipine,nifedipine, nicardipine, isradipine, nimodipine); the benzothiazepines(e.g., diltiazem), phenylalkylamines (e.g., verapamil); anddiarylaminopropylamine ether (e.g., bepridil).

[0011] The dihydropyridines, benzothiazepines and phenylalkylamines bindto distinct, but functionally coupled, sites on the α₁ subunit of L-typechannels at the interface of the IIIS6 and IVS6 transmembrane segments,such that the binding of any one class of drug can allostericallymodulate the binding of drugs in the other two classes and the highaffinity Ca⁺⁺ binding site in the channel (see GH Hockerman et al, Annu.Rev. Pharmacol. Toxicol. 37: 361-96 (1997)). It has been suggested thatmore than one high affinity binding site may exist for dihydropyridinesin voltage-dependent calcium channels (Kokubun et al, Molec. Pharmacol.30: 571-584 (1986)). However, studies reported in the scientificliterature cast doubt on this hypothesis. In particular, the antagonistactivities of a series of 1,n-alkanediylbis (1,4-dihydropyridines) wasreported to be essentially independent of the bridging carbon chainlength, and similar to that of the monomeric drugs (Joslyn et al., J.Med. Chem. 31: 1489-1492 (1988)).

[0012] The clinical shortcomings of drugs in current usage areconsiderable. Various benzothiazepines and phenylalkylamines, forexample, weaken cardiac contractility and are therefore contraindicatedin patients with left ventricular dysfunction. Other Ca⁺⁺ channelantagonists cause AV block, reflex tachycardia, excessive vasodilationand gastrointestinal problems. Their most common adverse side effectsinclude headache, flushing, hypotension, nausea, dizziness, fatigue,edema, abdominal pain, constipation, and the like. With few exceptions,the currently used drugs have a short duration of action and must beadministered frequently for sustained effects.

[0013] Thus, there continues to exist a need for novel compounds withgreater tissue selectivity, increased efficacy, reduced side effects anda more favorable duration of action.

SUMMARY OF THE INVENTION

[0014] This invention is directed to novel multibinding compounds thatbind to Ca⁺⁺ channels in mammalian tissues and can be used to treatdiseases and conditions mediated by such channels.

[0015] Accordingly, in one of its composition aspects, this invention isdirected to a multibinding compound and salts thereof comprising 2 to 10ligands which may be the same or different and which are covalentlyattached to a linker or linkers, which may be the same or different,each of said ligands comprising a ligand domain capable of binding to aCa⁺⁺ channel.

[0016] The multibinding compounds of this invention are preferablyrepresented by Formula I:

(L)_(p)(X)_(q)  I

[0017] where each L is a ligand that may be the same or different ateach occurrence;

[0018] X is a linker that may be the same or different at eachoccurrence;

[0019] p is an integer of from 2 to 10; and

[0020] q is an integer of from 1 to 20;

[0021] wherein each of said ligands comprises a ligand domain capable ofbinding to a Ca⁺⁺ channel.

[0022] Preferably q is less than p.

[0023] More preferably the linker is represented by the followingformula:

—X′—Z—(Y′—Z)_(m)—Y″—Z—X′—

[0024] in which:

[0025] m is an integer of from 0 to 20;

[0026] X′ at each separate occurrence is —O—, —S—, —S(O)—, —S(O)₂—,—NR—, —N⁺RR′—, —C(O)—, —C(O)O—, —C(O)NH—, —C(S), —C(S)O—, —C(S)NH— or acovalent bond, where R and R at each separate occurrence are as definedbelow for R′ and R″;

[0027] Z is at each separate occurrence selected from alkylene,substituted alkylene, alkylalkoxy, cycloalkylene, substitutedcycloalkylene, alkenylene, substituted alkenylene, alkynylene,substituted alkynylene, cycloalkenylene, substituted alkenylene,arylene, substituted arylene, heteroarylene, heterocyclene, substitutedheterocyclene, crown compounds, or a covalent bond;

[0028] Y′ and Y′ at each separate occurrence are selected from —S—S— ora covalent bond;

[0029] in which:

[0030] n is 0, 1 or 2; and

[0031] R′ and R″ at each separate occurrence are selected from hydrogen,alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, aryl, heteroaryl orheterocyclic.

[0032] Even more preferably, the ligands are selected from Ca⁺⁺ channelmodulators such as A-53930A, AE-0047, AGN-190604, AGN-190744, AH—1058,AHR-12742, AHR-16303B, AHR-16462B, AIT-110, AIT-111, AJ-3941, AM-336,amlopidine (including S-(−), R-(+), and racemic), anipamil, AP-1067,aranidipine, atosiban, azelnidipine, barnidipine, Bay-t-7207,Bay-y-5959, Bay-z-4406, BBR-2160, belfosdil, BIII-890-CL, bisaramil,BMS-181102, BMS-188107, BMY-43011, BRL-32872, buflomedil, CD-349,CD-832, CERM-12816, CGP-28932, cilnidipine, clentiazem, clevidipine,CNS-1067, CNS-1237, CNS-2103, CP-060S, CPC-301, CPC-317, CPU-86017,D-2024, darodipine, DHP-218, diltiazem, diperdipine, dopropidil,dotarazine, dronedarone, DTZ-323, E-047/1, efonidipine, EGIS-7229,elgodipine, emopamil, etomoxir, F-0401, fantofarone, fasudil, FCE-24265,FCE-26262, FCE-27335, FCE-27892, FCE-28718, felodipine, FPL-64176,FR-172516, FRG-8701, furnidipine, GS-386, iganidipine, ipenoxazone,isradipine, JTV-591, KP-840, KT-362, L-366682, lacidipine, LAS-0538,LCB-2514, lemildipine, lercanidipine, leualacin, lifarizine, LOE-908,lomerizine, lubeluzole, LY-042826, manidipine, McN-6186, mibefradil,monatepil , MR-14134, N-3601, NCC-1048, nefiracetam, nexopamil,nifedipine, nifedipine, Nifelan, nilvadipine, nimodipine, NNC-09-0026,NPS-568, NS-638, NS-649, NS-696, NS-7, OPC-8490, Org-13061, Org-30029,oxodipine, P-5, palonidipine, PCA-50922, PCA-50938, PCA-50941,PD-029361, PD-157667, PD-158143, PD-176078, pranidipine, QX-314,ranolazine, RHG-2716, RingCap, Ro-1 1-2933, RS-5773, RU-43945,RWJ-22108, RWJ-22726, RWJ-29009, RWJ-37868, S-12968, S-2150, S-312-d,SANK-71996, SB-201823, SB-206284A, SB-23736, SD-3212, semotiadil,SIB-1281, siratiazem, SKF-45675, SKF-96365, SKT-M-26, SL-34.0829,SL-87.0495, SM-6586, SNX-124, SNX-236, SNX-239, SNX-325, SNX-482,SQ-31727, SQ-33351, SQ-34399, SR-33805, TA-993, tamolarizine, TDN-345,temiverine, terodiline, TH9229, TN-871, U-88999, U-92032, U-92798,UCL-1439, UK-1656, UK-55444, UK-56593, UK-84149, verapamil, Verelan,vexibinol, VUF-8929, WAY-141520, XB-513, XT-044, Y-22516, YH-334,YM-1615-4, YM-430, Z-6568, zatebradine, ziconotide, and ZM-224832.

[0033] Particularly preferred Ca⁺⁺ channel modulators include verapamil,diltiazem, benziazem clentiazem, nicardipine, nifedipine, nilvadipine,nitredipine, nimodipine, isradipine, lacidipine, amlodipine,nisoldipine, isradipine, mibefradil, amlodipine, felodipine, nimodipine,bepridil, SQ 32,910 and SQ 32,428.

[0034] In a second embodiment, this invention is directed to apharmaceutical composition comprising a pharmaceutically acceptableexcipient and a therapeutically effective amount of one or moremultibinding compounds (or pharmaceutically acceptable salts thereof)comprising 2 to 10 ligands which may be the same or different and whichare covalently attached to a linker or linkers, which may be the same ordifferent, each of said ligands comprising a ligand domain capable ofbinding to a Ca⁺⁺ channel of a cell mediating mammalian diseases orconditions, thereby modulating the diseases or conditions.

[0035] In a third embodiment, this invention is directed to apharmaceutical composition comprising a pharmaceutically acceptableexcipient and a therapeutically effective amount of one or moremultibinding compounds represented by Formula I,

(L)_(p)(X)_(q)  I

[0036] or pharmaceutically acceptable salts thereof,

[0037] where each L is a ligand that may be the same or different ateach occurrence;

[0038] X is a linker that may be the same or different at eachoccurrence;

[0039] p is an integer of from 2 to 10; and

[0040] q is an integer of from 1 to 20;

[0041] wherein each of said ligands comprises a ligand domain capable ofbinding to a Ca⁺⁺ channel of a cell mediating mammalian diseases orconditions, thereby modulating the diseases or conditions. Preferably qis less than p.

[0042] In a fourth embodiment, this invention is directed to a methodfor modulating the activity of a Ca⁺⁺ channel in a biologic tissue,which method comprises contacting a tissue having a Ca⁺⁺ channel with amultibinding compound (or pharmaceutically acceptable salts thereof)under conditions sufficient to produce a change in the activity of thechannel in said tissue, wherein the multibinding compound comprises 2 to10 ligands which may be the same or different and which are covalentlyattached to a linker or linkers, which may be the same or different,each of said ligands comprising a ligand domain capable of binding to aCa⁺⁺ channel.

[0043] In a fifth embodiment, this invention is directed to a method fortreating a disease or condition in a mammal resulting from an activityof a Ca⁺⁺ channel, which method comprises administering to said mammal atherapeutically effective amount of a pharmaceutical compositioncomprising a pharmaceutically acceptable excipient and one or moremultibinding compounds (or pharmaceutically acceptable salts thereof)comprising 2 to 10 ligands which may be the same or different and whichare covalently attached to a linker or linkers, which may be the same ordifferent, each of said ligands comprising a ligand domain capable ofbinding to a Ca⁺⁺ channel of a cell mediating mammalian diseases orconditions.

[0044] In a sixth embodiment, this invention is directed to a method fortreating a disease or condition in a mammal resulting from an activityof a Ca⁺⁺ channel, which method comprises administering to said mammal atherapeutically effective amount of a pharmaceutical compositioncomprising a pharmaceutically acceptable excipient and one or moremultibinding compounds represented by Formula I,

(L)_(p)(X)_(q)  I

[0045] and pharmaceutically acceptable salts thereof,

[0046] where each L is a ligand that may be the same or different ateach occurrence;

[0047] X is a linker that may be the same or different at eachoccurrence;

[0048] p is an integer of from 2 to 10; and

[0049] q is an integer of from 1 to 20;

[0050] wherein each of said ligands comprises a ligand domain capable ofbinding to a Ca⁺⁺ channel of a cell mediating mammalian diseases orconditions.

[0051] Preferably q is less than p.

[0052] In a seventh embodiment, this invention relates to processes forpreparing the multibinding agents of Formula I.

[0053] In an eighth aspect, this invention is directed to generalsynthetic methods for generating large libraries of diverse multimericcompounds which multimeric compounds are candidates for possessingmultibinding properties. The diverse multimeric compound librariesprovided by this invention are synthesized by combining a linker orlinkers with a ligand or ligands to provide for a library of multimericcompounds wherein the linker and ligand each have complementaryfunctional groups permitting covalent linkage. The library of linkers ispreferably selected to have diverse properties such as valency, linkerlength, linker geometry and rigidity, hydrophilicity or hydrophobicity,amphiphilicity, acidity, basicity and polarization. The library ofligands is preferably selected to have diverse attachment points on thesame ligand, different functional groups at the same site of otherwisethe same ligand, and the like.

[0054] This invention is also directed to libraries of diversemultimeric compounds which multimeric compounds are candidates forpossessing multibinding properties. These libraries are prepared via themethods described above and permit the rapid and efficient evaluation ofwhat molecular constraints impart multibinding properties to a ligand ora class of ligands targeting a receptor.

[0055] Accordingly, in one of its method aspects, this invention isdirected to a method for identifying multimeric ligand compoundspossessing multibinding properties which method comprises:

[0056] (a) identifying a ligand or a mixture of ligands wherein eachligand contains at least one reactive functionality;

[0057] (b) identifying a library of linkers wherein each linker in saidlibrary comprises at least two functional groups having complementaryreactivity to at least one of the reactive functional groups of theligand;

[0058] (c) preparing a multimeric ligand compound library by combiningat least two stoichiometric equivalents of the ligand or mixture ofligands identified in (a) with the library of linkers identified in (b)under conditions wherein the complementary functional groups react toform a covalent linkage between said linker and at least two of saidligands; and

[0059] (d) assaying the multimeric ligand compounds produced in (c)above to identify multimeric ligand compounds possessing multibindingproperties.

[0060] In another of its method aspects, this invention is directed to amethod for identifying multimeric ligand compounds possessingmultibinding properties which method comprises:

[0061] (a) identifying a library of ligands wherein each ligand containsat least one reactive functionality;

[0062] (b) identifying a linker or mixture of linkers wherein eachlinker comprises at least two functional groups having complementaryreactivity to at least one of the reactive functional groups of theligand;

[0063] (c) preparing a multimeric ligand compound library by combiningat least two stoichiometric equivalents of the library of ligandsidentified in (a) with the linker or mixture of linkers identified in(b) under conditions wherein the complementary functional groups reactto form a covalent linkage between said linker and at least two of saidligands; and

[0064] (d) assaying the multimeric ligand compounds produced in (c)above to identify multimeric ligand compounds possessing multibindingproperties.

[0065] The preparation of the multimeric ligand compound library isachieved by either the sequential or concurrent combination of the twoor more stoichiometric equivalents of the ligands identified in (a) withthe linkers identified in (b). Sequential addition is preferred when amixture of different ligands is employed to ensure heterodimeric ormultimeric compounds are prepared. Concurrent addition of the ligandsoccurs when at least a portion of the multimer comounds prepared arehomomultimeric compounds.

[0066] The assay protocols recited in (d) can be conducted on themultimeric ligand compound library produced in (c) above, or preferably,each member of the library is isolated by preparative liquidchromatography mass spectrometry (LCMS).

[0067] In one of its composition aspects, this invention is directed toa library of multimeric ligand compounds which may possess multivalentproperties which library is prepared by the method comprising:

[0068] (a) identifying a ligand or a mixture of ligands wherein eachligand contains at least one reactive functionality;

[0069] (b) identifying a library of linkers wherein each linker in saidlibrary comprises at least two functional groups having complementaryreactivity to at least one of the reactive functional groups of theligand; and

[0070] (c) preparing a multimeric ligand compound library by combiningat least two stoichiometric equivalents of the ligand or mixture ofligands identified in (a) with the library of linkers identified in (b)under conditions wherein the complementary functional groups react toform a covalent linkage between said linker and at least two of saidligands.

[0071] In another of its composition aspects, this invention is directedto a library of multimeric ligand compounds which may possessmultivalent properties which library is prepared by the methodcomprising:

[0072] (a) identifying a library of ligands wherein each ligand containsat least one reactive functionality;

[0073] (b) identifying a linker or mixture of linkers wherein eachlinker comprises at least two functional groups having complementaryreactivity to at least one of the reactive functional groups of theligand; and

[0074] (c) preparing a multimeric ligand compound library by combiningat least two stoichiometric equivalents of the library of ligandsidentified in (a) with the linker or mixture of linkers identified in(b) under conditions wherein the complementary functional groups reactto form a covalent linkage between said linker and at least two of saidligands.

[0075] In a preferred embodiment, the library of linkers employed ineither the methods or the library aspects of this invention is selectedfrom the group comprising flexible linkers, rigid linkers, hydrophobiclinkers, hydrophilic linkers, linkers of different geometry, acidiclinkers, basic linkers, linkers of different polarization andamphiphilic linkers. For example, in one embodiment, each of the linkersin the linker library may comprise linkers of different chain lengthand/or having different complementary reactive groups. Such linkerlengths can preferably range from about 2 to 100 A.

[0076] In another preferred embodiment, the ligand or mixture of ligandsis selected to have reactive functionality at different sites on saidligands in order to provide for a range of orientations of said ligandon said multimeric ligand compounds. Such reactive functionalityincludes, by way of example, carboxylic acids, carboxylic acid halides,carboxyl esters, amines, halides, isocyanates, vinyl unsaturation,ketones, aldehydes, thiols, alcohols, anhydrides, and precursorsthereof. It is understood, of course, that the reactive functionality onthe ligand is selected to be complementary to at least one of thereactive groups on the linker so that a covalent linkage can be formedbetween the linker and the ligand.

[0077] In other embodiments, the multimeric ligand compound is homomeric(i.e., each of the ligands is the same, although it may be attached atdifferent points) or heterodimeric (i.e., at least one of the ligands isdifferent from the other ligands).

[0078] In addition to the combinatorial methods described herein, thisinvention provides for an iterative process for rationally evaluatingwhat molecular constraints impart multibinding properties to a class ofmultimeric compounds or ligands targeting a receptor. Specifically, thismethod aspect is directed to a method for identifying multimeric ligandcompounds possessing multibinding properties which method comprises:

[0079] (a) preparing a first collection or iteration of multimericcompounds which is prepared by contacting at least two stoichiometricequivalents of the ligand or mixture of ligands which target a receptorwith a linker or mixture of linkers wherein said ligand or mixture ofligands comprises at least one reactive functionality and said linker ormixture of linkers comprises at least two functional groups havingcomplementary reactivity to at least one of the reactive functionalgroups of the ligand wherein said contacting is conducted underconditions wherein the complementary functional groups react to form acovalent linkage between said linker and at least two of said ligands;

[0080] (b) assaying said first collection or iteration of multimericcompounds to assess which if any of said multimeric compounds possessmultibinding properties;

[0081] (c) repeating the process of (a) and (b) above until at least onemultimeric compound is found to possess multibinding properties;

[0082] (d) evaluating what molecular constraints imparted multibindingproperties to the multimeric compound or compounds found in the firstiteration recited in (a)-(c) above;

[0083] (e) creating a second collection or iteration of multimericcompounds which elaborates upon the particular molecular constraintsimparting multibinding properties to the multimeric compound orcompounds found in said first iteration;

[0084] (f) evaluating what molecular constraints imparted enhancedmultibinding properties to the multimeric compound or compounds found inthe second collection or iteration recited in (e) above;

[0085] (g) optionally repeating steps (e) and (f) to further elaborateupon said molecular constraints.

[0086] Preferably, steps (e) and (e are repeated at least two times,more preferably at from 2-50 times, even more preferably from 3 to 50times, and still more preferably at least 5-50 times.

BRIEF DESCRIPTION OF THE DRAWINGS

[0087]FIG. 1 is a highly schematic illustration of the transmembraneorganization of the α1 subunit of the voltage-gated Ca⁺⁺ channel.

[0088]FIG. 2 illustrates a method for optimizing the linker geometry forpresentation of ligands (filled circles) in bivalent compounds:

[0089] A. phenyldiacetylene core structure

[0090] B. cyclohexane dicarboxylic acid core structure

[0091]FIG. 3 shows exemplary linker “core” structures.

[0092] FIGS. 4-20 illustrate convenient methods for preparing themultibinding compounds of this invention.

[0093]FIG. 5 is a table of 741 calcium channel antagonists according tothe present invention.

DETAILED DESCRIPTION OF THE INVENTION

[0094] Biological systems in general are controlled by molecularinteractions between bioactive ligands and their receptors, in which thereceptor “recognizes” a molecule or a portion thereof (i.e., a liganddomain) to produce a biological effect. The voltage-gated Ca⁺⁺ channelsare considered to be pharmacological receptors: they possess specificbinding sites for ligands having agonist and antagonist activities; thebinding of ligands to such sites allosterically modulates Ca⁺⁺ fluxthrough the channel; the channel properties (i.e., gating and ionselectivity) are regulatable; and various channels are known toassociate with G-proteins (D. Rampe and D. J. Triggle, Prog. Drug Res.40: 191-238 (1993). Accordingly, diseases or conditions that involve, orare mediated by, Ca⁺⁺ channels can be treated with pharmacologicallyactive ligands that interact with such channels to initiate, modulate orabrogate transporter activity .

[0095] The interaction of a Ca⁺⁺ channel and a Ca⁺⁺ channel-bindingligand may be described in terms of “affinity” and “specificity”. The“affinity” and “specificity” of any given ligand-Ca⁺⁺ channelinteraction is dependent upon the complementarity of molecular bindingsurfaces and the energetic costs of complexation (i.e., the netdifference in free energy ΔG between bound and free states). Affinitymay be quantified by the equilibrium constant of complex formation, theratio of on/off rate constants, and/or by the free energy of complexformation. Specificity relates to the difference in binding affinity ofa ligand for different receptors.

[0096] The net free energy of interaction of such ligands with a Ca⁺⁺channel is the difference between energetic gains (enthalpy gainedthrough molecular complementarity and entropy gained through thehydrophobic effect) and energetic costs (enthalpy lost through decreasedsolvation and entropy lost through reduced translational, rotational andconformational degrees of freedom).

[0097] The compounds of this invention comprise 2 to 10 Ca⁺⁺channel-binding ligands covalently linked together and capable of actingas multibinding agents. Without wishing to be bound by theory, theenhanced activity of these compounds is believed to arise at least inpart from their ability to bind in a multivalent manner with multipleligand binding sites on a Ca⁺⁺ channel or channels, which gives rise toa more favorable net free energy of binding. Multivalent interactionsdiffer from collections of individual monovalent (univalent)interactions by being capable of providing enhanced biologic and/ortherapeutic effect. Multivalent binding can amplify binding affinitiesand differences in binding affinities, resulting in enhanced bindingspecificity as well as affinity.

[0098] Definitions

[0099] As used herein:

[0100] The term “alkyl” refers to a monoradical branched or unbranchedsaturated hydrocarbon chain, preferably having from 1 to 40 carbonatoms, preferably 1-10 carbon atoms, more preferably 1-6 carbon atoms,such as methyl, ethyl, n-propyl, isopropyl, n-butyl, secondary butyl,tert-butyl, n-hexyl, n-octyl, n-decyl, n-dodecyl, 2-ethyldodecyl,tetradecyl, and the like, unless otherwise indicated.

[0101] The term “substituted alkyl” refers to an alkyl group as definedabove having from 1 to 5 substituents selected from the group consistingof alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-aryl, —SO—heteroaryl,—SO₂-alkyl, —SO₂-aryl, —SO₂-heteroaryl, and —NRaRb, wherein R^(a) andR^(b) may be the same or different and and are chosen from hydrogen,optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl,alkynyl, aryl, heteroaryl and heterocyclic.

[0102] The term “alkylene” refers to a diradical of a branched orunbranched saturated hydrocarbon chain, preferably having from 1 to 40carbon atoms, preferably 1-10 carbon atoms, more preferably 1-6 carbonatoms. This term is exemplified by groups such as methylene (—CH₂—),ethylene (—CH₂CH₂—), the propylene isomers (e.g., —CH₂CH₂CH₂— and—CH(CH₃)CH₂—) and the like.

[0103] The term “substituted alkylene” refers to an alkylene group asdefined above having from 1 to 5 substituents selected from the groupconsisting of alkoxy, substituted alkoxy, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino,acyloxy, amino, aminoacyl, aminoacyloxy, oxyacylamino, azido, cyano,halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thiol,thioalkoxy, substituted thioalkoxy, aryl, aryloxy, thioaryloxy,heteroaryl, heteroaryloxy, thioheteroaryloxy, heterocyclic,heterocyclooxy, thioheterocyclooxy, nitro, and —NR^(a)R^(b), whereinR^(a) and R^(b) may be the same or different and are chosen fromhydrogen, optionally substituted alkyl, cycloalkyl, alkenyl,cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic. Additionally,such substituted alkylene groups include those where 2 substituents onthe alkylene group are fused to form one or more cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, heterocyclicor heteroaryl groups fused to the alkylene group. The term “substitutedalkylene” optionally includes an alkylene chain as defined above inwhich the carbon chain is interrupted by one or more atoms chosen fromO, S or N (e.g., ethers, sulfides and amines).

[0104] The term “alkaryl” or “aralkyl” refers to the groups-alkylene-aryl and -substituted alkylene-aryl in which alkylene and arylare as defined herein. Such alkaryl groups are exemplified by benzyl,phenethyl and the like.

[0105] The term “alkoxy” refers to the groups alkyl-O—, alkenyl-O—,cycloalkyl-O—, cycloalkenyl-O—, and alkynyl-O—, where alkyl, alkenyl,cycloalkyl, cycloalkenyl, and alkynyl are as defined herein. Preferredalkoxy groups are alkyl-O— and include, by way of example, methoxy,ethoxy, n-propoxy, iso -propoxy, n-butoxy, tert-butoxy, sec-butoxy,n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like

[0106] The term “substituted alkoxy” refers to the groups substitutedalkyl-O—, substituted alkenyl-O—, substituted cycloalkyl-O—, substitutedcycloalkenyl-O—, and substituted alkynyl-O— where substituted alkyl,substituted alkenyl, substituted cycloalkyl, substituted cycloalkenyland substituted alkynyl are as defined herein.

[0107] “Alkenyl” refers to a monoradical of a branched or unbranchedunsaturated hydrocarbon preferably having from 2 to 40 carbon atoms,preferably 2-10 carbon atoms, more preferably 2-6 carbon atoms, andpreferably having 1-6 double bonds. This term is further exemplified bysuch radicals as vinyl, prop-2-enyl, pent-3-enyl, hex-5-enyl,5-ethyldodec -3,6-dienyl, and the like.

[0108] The term “substituted alkenyl” refers to an alkenyl group asdefined above having from 1 to 5 substituents selected from the groupconsisting of alkoxy, substituted alkoxy, acyl, acylamino, acyloxy,amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen,hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thiol, thioalkoxy,substituted thioalkoxy, aryl, heteroaryl, heterocyclic, aryloxy,thioaryloxy, heteroaryloxy, thioheteroaryloxy, heterocyclooxy,thioheterocyclooxy, nitro, —SO-alkyl, —SO— substituted al kyl, —SO-aryl,—SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl,—SO₂-heteroaryl, and. —NR^(a)R^(b), wherein R^(a) and R^(b) may be thesame or different and are chosen from hydrogen, optionally substitutedalkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl andheterocyclic.

[0109] “Alkenylene” refers to a diradical of an unsaturated hydrocarbon,preferably having from 2 to 40 carbon atoms, preferably 2-10 carbonatoms, more preferably 2-6 carbon atoms, and preferably having 1-6double bonds. This term is further exemplified by such radicals as1,2-ethenyl, 1,3-prop-2-enyl, 1,5-pent-3-enyl, 1,4-hex-5-enyl,5-ethyl-1,12-dodec-3,6-dienyl, and the like.

[0110] The term “substituted alkenylene” refers to an alkenylene groupas defined above having from 1 to 5 substituents, selected from thegroup consisting of alkoxy, substituted alkoxy, acyl, acylamino,acyloxy, amino, aminoacyl, aminoacyloxy, oxyacylamino, azido, cyano,halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thiol,thioalkoxy, substituted thioalkoxy, aryl, aryloxy, thioaryloxy,heteroaryl, heteroaryloxy, thioheteroaryloxy, heterocyclic,heterocyclooxy, thioheterocyclooxy, nitro, and NR^(a)R^(b), whereinR^(a) and R^(b) may be the same or different and are chosen fromhydrogen, optionally substituted alkyl, cycloalkyl, alkenyl,cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic. Additionally,such substituted alkenylene groups include those where 2 substituents onthe alkenylene group are fused to form one or more cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,heterocyclic or heteroaryl groups fused to the alkenylene group.

[0111] “Alkynyl” refers to a monoradical of an unsaturated hydrocarbon,preferably having from 2 to 40 carbon atoms, preferably 2-10 carbonatoms, more preferably 2-6 carbon atoms, and preferably having 1-6triple bonds. This term is further exemplified by such radicals asacetylenyl, prop-2-ynyl, pent-3-ynyl, hex-5-ynyl,5-ethyldodec-3,6-diynyl, and the like.

[0112] The term “substituted alkynyl” refers to an alkynyl group asdefined above having from 1 to 5 substituents, selected from the groupconsisting of alkoxy, substituted alkoxy, acyl, acylamino, acyloxy,amino, aminoacyl, aminoacyloxy, oxyacylamino, azido, cyano, halogen,hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thiol, thioalkoxy,substituted thioalkoxy, aryl, aryloxy, thioaryloxy, heteroaryl,heteroaryloxy, thioheteroaryloxy, heterocyclic, heterocyclooxy,thioheterocycloxy, nitro, —SO-alkyl, —SO— substituted alkyl, —SO-aryl,—SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl,—SO₂-heteroaryl, SO₂-heterocyclic, NR^(a)R^(b), wherein R^(a) and R^(b)may be the same or different and are chosen from hydrogen, optionallysubstituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl,heteroaryl and heterocyclic.

[0113] “Alkynylene” refers to a diradical of an unsaturated hydrocarbonradical, preferably having from 2 to 40 carbon atoms, preferably 2-10carbon atoms, more preferably 2-6 carbon atoms, and preferably having1-6 triple bonds. This term is further exemplified by such radicals as 1,3-prop-2-ynyl, 1,5-pent-3-ynyl, 1,4-hex-5-ynyl,5-ethyl-1,12-dodec-3,6-diynyl, and the like.

[0114] The term “acyl” refers to the groups —CHO, alkyl-C(O)—,substituted alkyl-C(O)—, cycloalkyl-C(O)—, substituted cycloalkyl-C(O)—,cycloalkenyl-C(O)—, substituted cycloalkenyl-C(O)—, aryl-C(O)—,heteroaryl-C(O)— and heterocyclic-C(O)— where alkyl, substituted alkyl,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic are as defined herein.

[0115] The term “acylamino” refers to the group —C(O)NRR where each R isindependently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl,heterocyclic or where both R groups are joined to form a heterocyclicgroup (e.g., morpholine) wherein alkyl, substituted alkyl, aryl,heteroaryl and heterocyclic are as defined herein.

[0116] The term “aminoacyl” refers to the group —NRC(O)R where each R isindependently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, orheterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl andheterocyclic are as defined herein.

[0117] The term “aminoacyloxy” refers to the group —NRC(O)OR where eachR is independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl,or heterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl andheterocyclic are as defined herein.

[0118] The term “acyloxy” refers to the groups alkyl-C(O)O—, substitutedalkyl-C(O)O—, cycloalkyl-C(O)O—, substituted cycloalkyl-C(O)O—,aryl-C(O)O—, heteroaryl-C(O)O—, and heterocyclic-C(O)O— wherein alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, heteroaryl,and heterocyclic are as defined herein.

[0119] The term “aryl” refers to an unsaturated aromatic carbocyclicgroup of from 6 to 20 carbon atoms having a single ring (e.g., phenyl)or multiple condensed (fused) rings (e.g., naphthyl or anthryl).

[0120] Unless otherwise constrained by the definition for the arylsubstituent, such aryl groups can optionally be substituted with from 1to 5 substituents selected from the group consisting of acyloxy,hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, substituted alkyl, substituted alkoxy, substitutedalkenyl, substituted alkynyl, substituted cycloalkyl, substitutedcycloalkenyl, amino, aminoacyl, acylamino, alkaryl, aryl, aryloxy,azido, carboxyl, carboxylalkyl, cyano, halo, nitro, heteroaryl,heteroaryloxy, heterocyclic, heterocyclooxy, aminoacyloxy, oxyacylamino,thioalkoxy, substituted thioalkoxy, thioaryloxy, thioheteroaryloxy,—SO-alkyl, —SO—substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl, —SO₂-heteroaryl, trihalomethyl,NR^(a)R^(b), wherein R^(a) and R^(b) may be the same or different andare chosen from hydrogen, optionally substituted alkyl, cycloalkyl,alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic.Preferred aryl substituents include alkyl, alkoxy, halo, cyano, nitro,trihalomethyl, and thioalkoxy.

[0121] The term “aryloxy” refers to the group aryl-O— wherein the arylgroup is as defined above including optionally substituted aryl groupsas also defined above.

[0122] The term “arylene” refers to a diradical derived from aryl orsubstituted aryl as defined above, and is exemplified by 1,2-phenylene,1,3-phenylene, 1,4-phenylene, 1,2-naphthylene and the like.

[0123] The term “carboxyalkyl” refers to the group “—C(O)Oalkyl” wherealkyl is as defined above.

[0124] The term “cycloalkyl” refers to cyclic alkyl groups of from 3 to20 carbon atoms having a single cyclic ring or multiple condensed rings.Such cycloalkyl groups include, by way of example, single ringstructures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, andthe like, or multiple ring structures such as adamantanyl, and the like.

[0125] The term “substituted cycloalkyl” refers to cycloalkyl groupshaving from 1 to 5 substituents selected from the group consisting ofalkoxy, substituted alkoxy, cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, acyl, acylamino, acyloxy, amino, aminoacyl, aminoacyloxy,oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl,carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy,thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino,nitro, —SO-alkyl, —SO— substituted alkyl, —SO-aryl, —SO-heteroaryl,—SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl, —SO₂-heteroaryl, andNR^(a)R^(b), wherein R^(a) and R^(b) may be the same or different andare chosen from hydrogen, optionally substituted alkyl, cycloalkyl,alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic.

[0126] The term “cycloalkenyl” refers to cyclic alkenyl groups of from 4to 20 carbon atoms having a single cyclic ring or fused rings and atleast one point of internal unsaturation. Examples of suitablecycloalkenyl groups include, for instance, cyclobut-2-enyl,cyclopent-3-enyl, cyclooct-3-enyl and the like.

[0127] The term “substituted cycloalkenyl” refers to cycloalkenyl groupshaving from 1 to 5 substituents selected from the group consisting ofalkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO—substituted alkyl,—SO-aryl, —SO—heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl,—SO₂-heteroaryl, and NR^(a)R^(b), wherein R^(a) and R^(b) may be thesame or different and are chosen from hydrogen, optionally substitutedalkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl andheterocyclic.

[0128] The term “halo” or “halogen” refers to fluoro, chloro, bromo andiodo.

[0129] The term “heteroaryl” refers to an aromatic group of from 1 to 15carbon atoms and 1 to 4 heteroatoms selected from oxygen, nitrogen andsulfur within at least one ring (if there is more than one ring).

[0130] Unless otherwise constrained by the definition for the heteroarylsubstituent, such heteroaryl groups can be optionally substituted with 1to 5 substituents selected from the group consisting of acyloxy,hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, substituted alkyl, substituted alkoxy, substitutedalkenyl, substituted alkynyl, substituted cycloalkyl, substitutedcycloalkenyl, amino, aminoacyl, acylamino, alkaryl, aryl, aryloxy,azido, carboxyl, carboxylalkyl, cyano, halo, nitro, heteroaryl,heteroaryloxy, heterocyclic, heterocyclooxy, aminoacyloxy, oxyacylamino,thioalkoxy, substituted thioalkoxy, thioaryloxy, thioheteroaryloxy,—SO-alkyl, —SO— substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl, —SO₂-heteroaryl, trihalomethyl,mono-and di-alkylamino, mono- and NR^(a)R^(b), wherein R^(a) and R^(b)may be the same or different and are chosen from hydrogen, optionallysubstituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl,heteroaryl and heterocyclic. Preferred heteroaryls nclude pyridyl,pyrrolyl and furyl.

[0131] The term “heteroaryloxy” refers to the group heteroaryl-O—.

[0132] The term “heteroarylene” refers to the diradical group derivedfrom heteroaryl or substituted heteroaryl as defined above, and isexemplified by the groups 2,6-pyridylene, 2,4-pyridiylene,1,2-quinolinylene, 1,8-quinolinylene, 1,4-benzofuranylene,2,5-pyridinylene, 1,3-morpholinylene, 2,5-indolenyl, and the like.

[0133] The term “heterocycle” or “heterocyclic” refers to a monoradicalsaturated or unsaturated group having a single ring or multiplecondensed rings, from 1 to 40 carbon atoms and from 1 to 10 heteroatoms, preferably 1 to 4 heteroatoms, selected from nitrogen, sulfur,phosphorus, and/or oxygen within the ring.

[0134] Unless otherwise constrained by the definition for theheterocyclic substituent, such heterocyclic groups can be optionallysubstituted with 1 to 5, and preferably 1 to 3 substituents, selectedfrom the group consisting of alkoxy, substituted alkoxy, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl,acylamino, acyloxy, amino, aminoacyl, aminoacyloxy, oxyaminoacyl, cyano,halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy,thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substitutedthioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclic,heterocyclooxy, hydroxyamino, alkoxyamino, nitro, —SO-alkyl,—SO—substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl, —SO₂-heteroaryl, and NR^(a)R^(b),wherein R^(a) and R^(b) may be the same or different and are chosen fromhydrogen, optionally substituted alkyl, cycloalkyl, alkenyl,cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic. Suchheterocyclic groups can have a single ring or multiple condensed rings.

[0135] Examples of nitrogen heterocycles and heteroaryls include, butare not limited to, pyrrole, imidazole, pyrazole, pyridine, pyrazine,pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine,quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine,quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline,phenanthridine, acridine, phenanthroline, isothiazole, phenazine,isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline,piperidine, piperazine, indoline, morpholino, piperidinyl,tetrahydrofuranyl, and the like as well as N-alkoxy-nitrogen containingheterocycles.

[0136] A preferred class of heterocyclics include “crown compounds”which refers to a specific class of heterocyclic compounds having one ormore repeating units of the formula [—(CH₂—)_(m)Y—] where m is equal toor greater than 2, and Y at each separate occurrence can be O, N, S orP. Examples of crown compounds include, by way of example only,[—(CH₂)₃—NH—]₃, [—((CH₂)₂—O)₄—((CH₂)₂—NH)₂] and the like. Typically suchcrown compounds can have from 4 to 10 heteroatoms and 8 to 40 carbonatoms.

[0137] The term “heterocyclooxy” refers to the group heterocyclic-O—.

[0138] The term “thioheterocyclooxy” refers to the groupheterocyclic-S—.

[0139] The term “heterocyclene” refers to the diradical group derivedfrom a heterocycle as defined herein, and is exemplified by the groups2,6-morpholino, 2,5-morpholino and the like.

[0140] The term “oxyacylamino” refers to the group —OC(O)NRR where eachR is independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl,or heterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl andheterocyclic are as defined herein.

[0141] The term “thiol” refers to the group —SH.

[0142] The term “thioalkoxy” refers to the group —S-alkyl.

[0143] The term “substituted thioalkoxy” refers to the group—S—substituted alkyl.

[0144] The term “thioaryloxy” refers to the group aryl-S— wherein thearyl group is as defined above including optionally substituted arylgroups also defined above.

[0145] The term “thioheteroaryloxy” refers to the group heteroaryl-S—wherein the heteroaryl group is as defined above including optionallysubstituted aryl groups as also defined above.

[0146] As to any of the above groups which contain one or moresubstituents, it is understood, of course, that such groups do notcontain any substitution or substitution patterns which are stericallyimpractical and/or synthetically non-feasible. In addition, thecompounds of this invention include all stereochemical isomers arisingfrom the substitution of these compounds.

[0147] “Alkyl optionally interrupted by 1-5 atoms chosen from O, S, orN” refers to alkyl as defined above in which the carbon chain isinterrupted by O, S, or N. Within the scope are ethers, sulfides, andamines, for example 1-methoxydecyl, 1-pentyloxynonane,1-(2-isopropoxyethoxy)-4-methyinonane, 1-(2-ethoxyethoxy)dodecyl,2-(t-butoxy)heptyl, 1-pentylsulfanylnonane, nonylpentylamine, and thelike.

[0148] “Heteroarylalkyl” refers to heteroaryl as defined above linked toalkyl as defined above, for example pyrid-2-ylmethyl,8-quinolinylpropyl, and the like.

[0149] “Optional” or “optionally” means that the subsequently describedevent or circumstance may or may not occur, and that the descriptionincludes instances where said event or circumstance occurs and instancesin which it does not. For example, optionally substituted alkyl meansthat alkyl may or may not be substituted by those groups enumerated inthe definition of substituted alkyl.

[0150] The term “pharmaceutically acceptable salt” refers to salts whichretain the biological effectiveness and properties of the multibindingcompounds of this invention and which are not biologically or otherwiseundesirable. In many cases, the multibinding compounds of this inventionare capable of forming acid and/or base salts by virtue of the presenceof amino and/or carboxyl groups or groups similar thereto.

[0151] Pharmaceutically acceptable base addition salts can be preparedfrom inorganic and organic bases. Salts derived from inorganic bases,include by way of example only, sodium, potassium, lithium, ammonium,calcium and magnesium salts. Salts derived from organic bases include,but are not limited to, salts of primary, secondary and tertiary amines,such as alkyl amines, dialkyl amines, trialkyl amines, substituted alkylamines, di(substituted alkyl) amines, tri(substituted alkyl) amines,alkenyl amines, dialkenyl amines, trialkenyl amines, substituted alkenylamines, di(substituted alkenyl) amines, tri(substituted alkenyl) amines,cycloalkyl amines, di(cycloalkyl) amines, tri(cycloalkyl) amines,substituted cycloalkyl amines, disubstituted cycloalkyl amine,trisubstituted cycloalkyl amines, cycloalkenyl amines, di(cycloalkenyl)amines, tri(cycloalkenyl) amines, substituted cycloalkenyl amines,disubstituted cycloalkenyl amine, trisubstituted cycloalkenyl amines,aryl amines, diaryl amines, triaryl amines, heteroaryl amines,diheteroaryl amines, triheteroaryl amines, heterocyclic amines,diheterocyclic amines, triheterocyclic amines, mixed di— and tri- amineswhere at least two of the substituents on the amine are different andare selected from the group consisting of alkyl, substituted alkyl,alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl, heterocyclic,and the like. Also included are amines where the two or threesubstituents, together with the amino nitrogen, form a heterocyclic orheteroaryl group.

[0152] Examples of suitable amines include, by way of example only,isopropylamine, trimethyl amine, diethyl amine, tri(iso-propyl) amine,tri(n-propyl) amine, ethanolamine, 2-dimethylaminoethanol, tromethamine,lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline,betaine, ethylenediamine, glucosamine, N-alkylglucamines, theobromine,purines, piperazine, piperidine, morpholine, N-ethylpiperidine, and thelike. It should also be understood that other carboxylic acidderivatives would be useful in the practice of this invention, forexample, carboxylic acid amides, including carboxamides, lower alkylcarboxamides, dialkyl carboxamides, and the like.

[0153] Pharmaceutically acceptable acid addition salts may be preparedfrom inorganic and organic acids. Salts derived from inorganic acidsinclude hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like. Salts derived from organic acids includeacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid,malic acid, malonic acid, succinic acid, maleic acid, fumaric acid,tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluene-sulfonic acid,salicylic acid, and the like.

[0154] The term “library” refers to at least 3, preferably from 10² to10⁹ and more preferably from 10² to 10⁴ multimeric compounds.Preferably, these compounds are prepared as a multiplicity of compoundsin a single solution or reaction mixture which permits facile synthesisthereof. In one embodiment, the library of multimeric compounds can bedirectly assayed for multibinding properties. In another embodiment,each member of the library of multimeric compounds is first isolatedand, optionally, characterized. This member is then assayed formultibinding properties.

[0155] The term “collection” refers to a set of multimeric compoundswhich are prepared either sequentially or concurrently (e.g.,combinatorially). The collection comprises at least 2 members;preferably from 2 to 10⁹ members and still more preferably from 10 to10⁴ members.

[0156] The term “multimeric compound” refers to compounds comprisingfrom 2 to 10 ligands covalently connected through at least one linkerwhich compounds may or may not possess multibinding properties (asdefined herein).

[0157] The term “pseudohalide” refers to functional groups which reactin displacement reactions in a manner similar to a halogen. Suchfunctional groups include, by way of example, mesyl, tosyl, azido andcyano groups.

[0158] The term “protecting group” or “blocking group” refers to anygroup which when bound to one or more hydroxyl, thiol, amino or carboxylgroups of the compounds prevents reactions from occurring at thesegroups and which protecting group can be removed by conventionalchemical or enzymatic steps to reestablish the hydroxyl, thiol, amino orcarboxyl group. See, generally, T. W. Greene & P. G. M. Wuts “ProtectiveGroups in Organic Synthesis,” 2nd Ed, 1991, John Wiley and Sons, N.Y.

[0159] The particular removable blocking group employed is not criticaland preferred removable hydroxyl blocking groups include conventionalsubstituents such as allyl, benzyl, acetyl, chloroacetyl, thiobenzyl,benzylidine, phenacyl, t-butyl-diphenylsilyl and any other group thatcan be introduced chemically onto a hydroxyl functionality and laterselectively removed either by chemical or enzymatic methods in mildconditions compatible with the nature of the product.

[0160] Preferred removable amino blocking groups include conventionalsubstituents such as t-butyoxycarbonyl (t-BOC), benzyloxycarbonyl (CBZ),fluorenylmethoxycarbonyl (FMOC), allyloxycarbonyl (ALOC) and the like,which can be removed by conventional conditions compatible with thenature of the product.

[0161] Preferred carboxyl protecting groups include esters such asmethyl, ethyl, propyl, t-butyl etc. which can be removed by mildhydrolysis conditions compatible with the nature of the product.

[0162] As used herein, the terms “inert organic solvent” or “inertsolvent” mean a solvent inert under the conditions of the reaction beingdescribed in conjunction therewith [including, for example, benzene,toluene, acetonitrile, tetrahydrofuran (“THF”), dimethylformamide(“DMF”), chloroform (“CHCl₃”), methylene chloride (or dichloromethane or“CH₂Cl₂”), diethyl ether, ethyl acetate, acetone, methylethyl ketone,methanol, ethanol, propanol, isopropanol, tert-butanol, dioxane,pyridine, and the like]. Unless specified to the contrary, the solventsused in the reactions of the present invention are inert solvents.

[0163] The term “Ca⁺⁺ channel” refers to a structure comprised ofintegral membrane proteins that functions to allow Ca⁺⁺ to equilibrateacross a membrane according to its electrochemical gradient and at ratesthat are diffusion-limited. Examples of various types of Ca⁺⁺ channelsare given in Table 1 (Appendix).

[0164] “Ligand” as used herein denotes a compound that is a bindingpartner for a Ca⁺⁺ channel receptor, and is bound thereto, for example,by complementarity. The specific region or regions of the ligandmolecule that is recognized by the ligand binding site of a Ca⁺⁺ channelreceptor is designated as the “ligand domain”. A ligand may be eithercapable of binding to a receptor by itself, or may require the presenceof one or more non-ligand components for binding (e.g. ions, a lipidmolecule, a solvent molecule, and the like).

[0165] Ligands useful in this invention comprise Ca⁺⁺ channel modulatorssuch as verapamil (a phenylalkylamine), diltiazem (a benzothiazepine),nicardipine, nifedipine, isradipine, amlodipine, mibefradil, felodipineand nimodipine (dihydropyridines), and bepridil (adiarylaminopropylamine ether). See Tables 4 and 5, Appendix forstructures of various dihydropyridine, phenylalkylamine, andbenzothiazepine ligands.

[0166] While it is contemplated that many calcium channel ligands thatare currently known can be used in the preparation of multibindingcompounds of this invention (see Table 2 (Appendix)), it should beunderstood that portions of the ligand structure that are not essentialfor molecular recognition and binding activity (i.e., that are not partof the ligand domain) may be varied substantially, replaced withunrelated structures and, in some cases, omitted entirely withoutaffecting the binding interaction. Accordingly, it should be understoodthat the term “ligand” is not intended to be limited to compounds knownto be useful as Ca⁺⁺ channel receptor-binding compounds (e.g., knowndrugs), in that ligands that exhibit marginal activity or lack usefulactivity as monomers can be highly active as multibinding compounds,because of the biological benefit conferred by multivalency. The primaryrequirement for a ligand as defined herein is that it has a liganddomain, as defined above, which is available for binding to arecognition site on a Ca⁺⁺ channel.

[0167] For purposes of the present invention, the term “ligand” or“ligands” is intended to include the racemic ligands as well as theindividual stereoisomers of the ligands, including pure enantiomers andnon-racemic mixtures thereof. A large number of known calcium channelligands have at least one chiral center and show stereoselectivepharmacokinetics and pharmacologic activity (reviewed in Tokuma,Y andNoguchi, H., J. Chromatography A, 694: 181-193 (1995)). The scope of theinvention as described and claimed encompasses the racemic forms of theligands as well as the individual enantiomers and non-racemic mixturesthereof.

[0168] The term “ligand binding site” as used herein denotes a site on aCa⁺⁺channel receptor that recognizes a ligand domain and provides abinding partner for the ligand. The ligand binding site may be definedby monomeric or multimeric structures. This interaction may be capableof producing a unique biological effect, for example agonism,antagonism, modulation, or may maintain an ongoing biological event, andthe like.

[0169] It should be recognized that the ligand binding sites of Ca⁺⁺channel receptors that participate in biological multivalent bindinginteractions are constrained to varying degrees by their intra— andintermolecular associations. For example, Ca⁺⁺ channel ligand bindingsites may be covalently joined in a single structure, noncovalentlyassociated in one or more multimeric structures, embedded in a membraneor biopolymer matrix, and so on, and therefore have less translationaland rotational freedom than if the same sites were present as monomersin solution.

[0170] The terms “agonism” and “antagonism” are well known in the art.As used herein, the term “agonist” refers to a ligand that when bound toa Ca⁺⁺channel stimulates its activity. The term “antagonist” refers to aligand that when bound to a Ca⁺⁺ channel inhibits its activity. Channelblock or activation may result from allosteric effects of ligand bindingto the channel rather than occupancy of the channel pore. Theseallosteric effects may produce changes in protein conformation thataffect Ca⁺⁺ binding sites, gating mechanisms and/or the pore region(i.e., ion permeation).

[0171] A channel can exist in three modes: mode 0 (where the channel haszero probability of opening); mode 1 (where the channel opens frequentlybut transiently) and mode 2, where the channel remains open forrelatively long periods of time (Hess et al, Nature 311: 538-544(1984)). The probability that a channel will exist in one of these threestates changes with voltage. A given ligand may have different bindingaffinities for different states, and thereby be capable of producingagonist or antagonist activity.

[0172] The term “modulatory effect” is intended to refer to the abilityof a ligand to change the activity of a Ca⁺⁺ channel through binding tothe channel.

[0173] “Multibinding agent” or “multibinding compound” refers herein toa compound that has from 2 to 10 Ca⁺⁺ channel ligands as defined herein(which may be the same or different) covalently bound to one or morelinkers (which may be the same or different), and is capable ofmultivalency, as defined below.

[0174] A multibinding compound provides an improved biologic and/ortherapeutic effect compared to that of the same number of unlinkedligands available for binding to the ligand binding sites on a Ca⁺⁺channel or channels. Examples of improved “biologic and/or therapeuticeffect” include increased ligand-receptor binding interactions (e.g.,increased affinity, increased ability to elicit a functional change inthe target, improved kinetics), increased selectivity for the target,increased potency, increased efficacy, decreased toxicity, increasedtherapeutic index, improved duration of action, improvedbioavailability, improved pharmacokinetics, improved activity spectrum,, and the like. The multibinding compounds of this invention willexhibit at least one, and preferably more than one, of theabove-mentioned effects.

[0175] “Univalency” as used herein refers to a single bindinginteraction between one ligand with one ligand binding site as definedherein. It should be noted that a compound having multiple copies of aligand (or ligands) exhibits univalency when only one ligand of thatcompound interacts with a ligand binding site. Examples of univalentinteractions are depicted below.

[0176] “Multivalency” as used herein refers to the concurrent binding offrom 2 to 10 linked ligands, which may be the same or different, and twoor more corresponding ligand binding sites, which may be the same ordifferent. An example of trivalent binding is depicted below forillustrative purposes.

[0177] It should be understood that not all compounds that containmultiple copies of a ligand attached to a linker necessarily exhibit thephenomena of multivalency, i.e., that the biologic and/or therapeuticeffect of the multibinding agent is greater than that of the same numberof unlinked ligands made available for binding to the ligand bindingsites. For multivalency to occur, the ligand domains of the ligands thatare linked together must be presented to their cognate ligand bindingsites by the linker or linkers in a specific manner in order to bringabout the desired ligand-orienting result, and thus produce amultibinding interaction.

[0178] The term “linker”, identified where appropriate by the symbol X,refers to a group or groups that covalently links from 2 to 10 ligands(as defined above) in a manner that provides a multibinding compoundcapable of multivalency. The linker is a ligand-orienting entity thatpermits attachment of multiple copies of a ligand (which may be the sameor different) thereto.

[0179] The term “linker” includes everything that is not considered tobe part of the ligand, e.g., ancillary groups such as solubilizinggroups, lipophilic groups, groups that alter pharmacodynamics orpharmacokinetics, groups that modify the diffusability of themultibinding compound, spacers that attach the ligand to the linker,groups that aid the ligand-orienting function of the linker, forexample, by imparting flexibility or rigidity to the linker as a whole,or to a portion thereof, and so on. The term “linker” does not, however,cover solid inert supports such as beads, glass particles, rods, and thelike, but it is to be understood that the multibinding compounds of thisinvention can be attached to a solid support if desired, for example,for use in separation and purification processes and for similarapplications.

[0180] The extent to which the previously discussed enhanced activity ofmultibinding compounds is realized in this invention depends upon theefficiency with which the linker or linkers that joins the ligandspresents them to their array of ligand binding sites. Beyond presentingthese ligands for multivalent interactions with ligand binding sites,the linker spatially constrains these interactions to occur withindimensions defined by the linker.

[0181] The linkers used in this invention are selected to allowmultivalent binding of ligands to any desired ligand binding sites of aCa⁺⁺ channel, whether such sites are located interiorly (e.g., within achannel/translocation pore), both interiorly and on the periphery of achannel, at the boundary region between the lipid bilayer and thechannel, or at any intermediate position thereof. The preferred linkerlength will vary depending on the distance between adjacent ligandbinding sites, and the geometry, flexibility and composition of thelinker. The length of the linker will preferably be in the range ofabout 2 Å to about 100 Å, more preferably from about 2 Å to about 50 Åand even more preferably from about 3 Å to about 10 Å.

[0182] The ligands are covalently attached to the linker or linkersusing conventional chemical techniques. The reaction chemistriesresulting in such linkage are well known in the art and involve the useof reactive functional groups present on the linker and ligand.Preferably, the reactive functional groups on the linker are selectedrelative to the functional groups available on the ligand for coupling,or which can be introduced onto the ligand for this purpose. Again, suchreactive functional groups are well known in the art. For example,reaction between a carboxylic acid of either the linker or the ligandand a primary or secondary amine of the ligand or the linker in thepresence of suitable well-known activating agents results in formationof an amide bond covalently linking the ligand to the linker; reactionbetween an amine group of either the linker or the ligand and a sulfonylhalide of the ligand or the linker results in formation of a sulfonamidebond covalently linking the ligand to the linker; and reaction betweenan alcohol or phenol group of either the linker or the ligand and analkyl or aryl halide of the ligand or the linker results in formation ofan ether bond covalently linking the ligand to the linker. Table 3(Appendix) illustrates numerous reactive functional groups and theresulting bonds formed by reaction therebetween. Where functional groupsare lacking, they can be created by suitable chemistries that aredescribed in standard organic chemistry texts such as J. March,“Advanced Organic Chemistry”. 4^(th) Edition, (Wiley-lnterscience (NewYork), 1992.

[0183] The relative orientation in which the ligand domains aredisplayed depends both on the particular point or points of attachmentof the ligands to the linker, and on the framework geometry. Thedetermination of where acceptable substitutions can be made on a ligandis typically based on prior knowledge of structure-activityrelationships of the ligand and/or congeners and/or structuralinformation about ligand-receptor complexes (e.g., X-raycrystallography, NMR, and the like). Such positions and syntheticprotocols for linkage are well known in the art and can be determined bythose with ordinary skill in the art (see Methods of Preparation andFIGS. 4-20 of the Appendix). Following attachment of a ligand to thelinker or linkers, or to a significant portion thereof (e.g., 2-10 atomsof linker), the linker-ligand conjugate may be tested for retention ofactivity in a relevant assay system (see Utility and Testing below forrepresentative assays).

[0184] At present, it is preferred that the multibinding compound is abivalent compound in which two ligands are covalently linked, or atrivalent compound, in which three ligands are covalently linked. Linkerdesign is further discussed under Methods of Preparation.

[0185] “Potency” as used herein refers to the minimum concentration atwhich a ligand is able to achieve a desirable biological or therapeuticeffect. The potency of a ligand is typically proportional to itsaffinity for its receptor. In some cases, the potency may benon-linearly correlated with its affinity. In comparing the potency oftwo drugs, e.g., a multibinding agent and the aggregate of its unlinkedligand, the dose-response curve of each is determined under identicaltest conditions (e.g. in an in vitro or in vivo assay, in an appropriateanimal model). The finding that the multibinding agent produces anequivalent biologic or therapeutic effect at a lower concentration thanthe aggregate unlinked ligand (e.g. on a per weight, per mole or perligand basis) is indicative of enhanced potency.

[0186] “Selectivity” or “specificity” is a measure of the bindingpreferences of a ligand for different receptors. The selectivity of aligand with respect to its target receptor relative to another receptoris given by the ratio of the respective values of Kd (i.e., thedissociation constants for each ligand-receptor complex) or, in caseswhere a biological effect is observed below the K_(d), the ratio of therespective EC₅₀s or lC₅₀s (i.e., the concentrations that produce 50% ofthe maximum response for the ligand interacting with the two distinctreceptors).

[0187] The term “treatment” refers to any treatment of a disease orcondition in a mammal, particularly a human, and includes:

[0188] (i) preventing the disease or condition from occurring in asubject which may be predisposed to the condition but has not yet beendiagnosed with the condition and, accordingly, the treatment constitutesprophylactic treatment for the pathologic condition;

[0189] (ii) inhibiting the disease or condition, i.e., arresting itsdevelopment;

[0190] (iii) relieving the disease or condition, i.e., causingregression of the disease or condition; or

[0191] (iv) relieving the symptoms resulting from the disease orcondition without addressing the underlying disease or condition, e.g.,relieving symptoms of angina pectoris and other conditions of ischemiabut not an underlying cause such as, for example, atheroscleroticdisease or hypertension.

[0192] The phrase “disease or condition which is modulated by treatmentwith a multibinding Ca⁺⁺ channel ligand” covers all disease statesand/or conditions that are generally acknowledged in the art to beusefully treated with a ligand for a Ca⁺⁺ channel in general, and thosedisease states and/or conditions that have been found to be usefullytreated by a specific multibinding compound of our invention, i.e., thecompounds of Formula I. Such disease states include, by way of exampleonly, hypertension, angina pectoris (particularly vasospastic angina andunstable angina), cerebral ischemia, cardiac arrythmias (particularly,arrythmias resulting from calcium-related changes in membrane potentialand conduction), cardiac hypertrophy due to systolic or diastolicoverload, congestive heart failure, migraine, Raynaud's disease, acuterenal failure due to prolonged renal ischemia, and the like.

[0193] The term “therapeutically effective amount” refers to that amountof multibinding compound that is sufficient to effect treatment, asdefined above, when administered to a mammal in need of such treatment.The therapeutically effective amount will vary depending upon thesubject and disease condition being treated, the weight and age of thesubject, the severity of the disease condition, the manner ofadministration and the like, which can readily be determined by one ofordinary skill in the art.

[0194] The term “pharmaceutically acceptable excipient” is intended toinclude vehicles and carriers capable of being coadministered with amultibinding compound to facilitate the performance of its intendedfunction. The use of such media for pharmaceutically active substancesis well known in the art. Examples of such vehicles and carriers includesolutions, solvents, dispersion media, delay agents, emulsions and thelike. Any other conventional carrier suitable for use with themultibinding compounds also falls within the scope of the presentinvention.

METHODS OF PREPARATION

[0195] Linkers

[0196] The linker or linkers, when covalently attached to multiplecopies of the ligands, provides a biocompatible, substantiallynon-immunogenic multibinding compound. The biological activity of themultibinding Ca⁺⁺ channel compound is highly sensitive to the geometry,composition, size, length, flexibility or rigidity, the presence orabsence of anionic or cationic charge, the relative hydrophobicity/hydrophilicity, and similar properties of the linker. Accordingly, thelinker is preferably chosen to maximize the biological activity of thecompound. The linker may be biologically “neutral,” i.e., not itselfcontribute any additional biological activity to the multibindingcompound, or it may be chosen to further enhance the biological activityof the compound. In general, the linker may be chosen from any organicmolecule construct that orients two or more ligands for binding to thereceptors to permit multivalency. In this regard, the linker can beconsidered as a “framework” on which the ligands are arranged in orderto bring about the desired ligand-orienting result, and thus produce amultibinding compound.

[0197] For example, different orientations of ligands can be achieved byvarying the geometry of the framework (linker) by use of mono- orpolycyclic groups, such as aryl and/or heteroaryl groups, or structuresincorporating one or more carbon-carbon multiple bonds (alkenyl,alkenylene, alkynyl or alkynylene groups). The optimal geometry andcomposition of frameworks (linkers) used in the multibinding compoundsof this invention are based upon the properties of their intendedreceptors. For example, it is preferred to use rigid cyclic groups(e.g., aryl, heteroaryl), or non-rigid cyclic groups (e.g., cycloalkylor crown groups) to reduce conformational entropy when such may benecessary to achieve energetically coupled binding.

[0198] Different hydrophobic/hydrophilic characteristics of the linkeras well as the presence or absence of charged moieties can readily becontrolled by the skilled artisan. For example, the hydrophobic natureof a linker derived from hexamethylene diamine (H₂N(CH₂)₆NH₂) or relatedpolyamines can be modified to be substantially more hydrophilic byreplacing the alkylene group with a poly(oxyalkylene) group such asfound in the commercially available “Jeffamines” (class of surfactants).

[0199] Different frameworks can be designed to provide preferredorientations of the ligands. The identification of an appropriateframework geometry for ligand domain presentation is an important firststep in the construction of a multi binding agent with enhancedactivity. Systematic spatial searching strategies can be used to aid inthe identification of preferred frameworks through an iterative process.FIG. 2 (Appendix) illustrates a useful strategy for determining anoptimal framework display orientation for ligand domains and can be usedfor preparing the bivalent compounds of this invention. Variousalternative strategies known to those skilled in the art of moleculardesign can be substituted for the one described here.

[0200] As shown in FIG. 2, the ligands (shown as filled circles) areattached to a central core structure such as phenyidiacetylene (Panel A)or cyclohexane dicarboxylic acid (Panel B). The ligands are spaced apartfrom the core by an attaching moiety of variable lengths m and n. If theligand possesses multiple attachment sites (see discussion below), theorientation of the ligand on the attaching moiety may be varied as well.The positions of the display vectors around the central core structuresare varied, thereby generating a collection of compounds. Assay of eachof the individual compounds of a collection generated as described willlead to a subset of compounds with the desired enhanced activities(e.g., potency, selectivity). The analysis of this subset using atechnique such as Ensemble Molecular Dynamics will suggest a frameworkorientation that favors the properties desired.

[0201] The process may require the use of multiple copies of the samecentral core structure or combinations of different types of displaycores. It is to be noted that core structures other than those shownhere can be used for determining the optimal framework displayorientation of the ligands. The above-described technique can beextended to trivalent compounds and compounds of higher-order valency.

[0202] A wide variety of linkers is commercially available (see, e.g.,Chem Sources USA and Chem Sources International; the ACD electronicdatabase; and Chemical Abstracts). Many of the linkers that are suitablefor use in this invention fall into this category. Others can be readilysynthesized by methods known in the art, and as described below.Examples of linkers include aliphatic moieties, aromatic moieties,steroidal moieties, peptides, and the like. Specific examples arepeptides or polyamides, hydrocarbons, aromatics, heterocyclics, ethers,lipids, cationic or anionic groups, or a combination thereof.

[0203] Examples are given below and in FIG. 3 (Appendix), but it shouldbe understood that various changes may be made and equivalents may besubstituted without departing from the true spirit and scope of theinvention. For example, properties of the linker can be modified by theaddition or insertion of ancillary groups into the linker, for example,to change the solubility of the multibinding compound (in water, fats,lipids, biological fluids, etc.), hydrophobicity, hydrophilicity, linkerflexibility, antigenicity, stability, and the like. For example, theintroduction of one or more poly(ethylene glycol) (PEG) groups onto thelinker enhances the hydrophilicity and water solubility of themultibinding compound, increases both molecular weight and molecularsize and, depending on the nature of the unPEGylated linker, mayincrease the in vivo retention time. Further, PEG may decreaseantigenicity and potentially enhances the overall rigidity of thelinker.

[0204] Ancillary groups that enhance the water solubility/hydrophilicityof the linker, and accordingly, the resulting multibinding compounds,are useful in practicing this invention. Thus, it is within the scope ofthe present invention to use ancillary groups such as, for example,small repeating units of ethylene glycols, alcohols, polyols, (e.g.,glycerin, glycerol propoxylate, saccharides, including mono-,oligosaccharides, etc.) carboxylates (e.g., small repeating units ofglutamic acid, acrylic acid, etc.), amines (e.g.,tetraethylenepentamine), and the like to enhance the water solubilityand/or hydrophilicity of the multibinding compounds of this invention.In preferred embodiments, the ancillary group used to improve watersolubility I hydrophilicity will be a polyether. In particularlypreferred embodiments, the ancillary group will contain a small numberof repeating ethylene oxide (—CH₂CH₂O—) units.

[0205] The incorporation of lipophilic ancillary groups within thestructure of the linker to enhance the lipophilicity and/orhydrophobicity of the compounds of Formula I is also within the scope ofthis invention. Lipophilic groups useful with the linkers of thisinvention include, but are not limited to, lower alkyl, aromatic groupsand polycyclic aromatic groups. The aromatic groups may be eitherunsubstituted or substituted with other groups, but are at leastsubstituted with a group which allows their covalent attachment to thelinker. As used herein the term “aromatic groups” incorporates botharomatic hydrocarbons and heterocyclic aromatics. Other lipophilicgroups useful with the linkers of this invention include fatty acidderivatives which may or may not form micelles in aqueous medium andother specific lipophilic groups which modulate interactions between themultibinding compound and biological membranes.

[0206] Also within the scope of this invention is the use of ancillarygroups which result in the compound of Formula I being incorporated intoa vesicle, such as a liposome, or a micelle. The term “lipid” refers toany fatty acid derivative that is capable of forming a bilayer ormicelle such that a hydrophobic portion of the lipid material orientstoward the bilayer while a hydrophilic portion orients toward theaqueous phase. Hydrophilic characteristics derive from the presence ofphosphate, carboxylic, sulfato, amino, sulfhydryl, nitro and other likegroups well known in the art. Hydrophobicity could be conferred by theinclusion of groups that include, but are not limited to, long chainsaturated and unsaturated aliphatic hydrocarbon groups of up to 20carbon atoms and such groups substituted by one or more aryl,heteroaryl, cycloalkyl, and/or heterocyclic group(s). Preferred lipidsare phosphoglycerides and sphingolipids, representative examples ofwhich include phosphatidylcholine, phosphatidylethanolamine,phosphatidylserine, phosphatidylinositol, phosphatidic acid,palmitoyleoyl phosphatidylcholine, lysophosphatidylcholine,lysophosphatidyl-ethanolamine, dipalmitoylphosphatidylcholine,dioleoylphosphatidylcholine, distearoyl-phosphatidylcholine anddilinoleoylphosphatidylcholine. Other compounds lacking phosphorus, suchas sphingolipid and glycosphingolipid families, are also within thegroup designated as lipid. Additionally, the amphipathic lipidsdescribed above may be mixed with other lipids including triglyceridesand sterols.

[0207] The flexibility of the linker can be manipulated by the inclusionof ancillary groups which are bulky and/or rigid. The presence of bulkyor rigid groups can hinder free rotation about bonds in the linker, orbonds between the linker and the ancillary group(s), or bonds betweenthe linker and the functional groups. Rigid groups can include, forexample, those groups whose conformational freedom is restrained by thepresence of rings and/or π-bonds, for example, aryl, heteroaryl andheterocyclic groups. Other groups which can impart rigidity includepolypeptide groups such as oligo- or polyproline chains.

[0208] Rigidity can also be imparted electrostatically. Thus, if theancillary groups are either positively or negatively charged, thesimilarly charged ancillary groups will force the linker into aconfiguration affording the maximum distance between each of the likecharges. The energetic cost of bringing the like-charged groups closerto each other, which is inversely related to the square of the distancebetween the groups, will tend to hold the linker in a configuration thatmaintains the separation between the like-charged ancillary groups.Further, ancillary groups bearing opposite charges will tend to beattracted to their oppositely charged counterparts and potentially mayenter into both inter- and intramolecular ionic bonds. This non-covalentmechanism will tend to hold the linker in a conformation which allowsbonding between the oppositely charged groups. The addition of ancillarygroups which are charged, or alternatively, protected groups that bear alatent charge which is unmasked, following addition to the linker, bydeprotection, a change in pH, oxidation, reduction or other mechanismsknown to those skilled in the art, is within the scope of thisinvention. Bulky groups can include, for example, large atoms, ions(e.g., iodine, sulfur, metal ions, etc.) or groups containing largeatoms, polycyclic groups, including aromatic groups, non-aromatic groupsand structures incorporating one or more carbon-carbon n-bonds (i.e.,alkenes and alkynes). Bulky groups can also include oligomers andpolymers which are branched- or straight-chain species. Species that arebranched are expected to increase the rigidity of the structure more perunit molecular weight gain than are straight-chain species.

[0209] In preferred embodiments, rigidity (entropic control) is impartedby the presence of alicyclic (e.g., cycloalkyl), aromatic andheterocyclic groups. In other preferred embodiments, this comprises oneor more six-membered rings. In still further preferred embodiments, thering is an aryl group such as, for example, phenyl or naphthyl, or amacrocyclic ring such as, for example, a crown compound.

[0210] In view of the above, it is apparent that the appropriateselection of a linker group providing suitable orientation, entropy andphysico-chemical properties is well within the skill of the art.

[0211] Eliminating or reducing antigenicity of the multibindingcompounds described herein is also within the scope of this invention.In certain cases, the antigenicity of a multibinding compound may beeliminated or reduced by use of groups such as, for example,poly(ethylene glycol).

[0212] Compounds of Formula I

[0213] As explained above, the multibinding compounds described hereincomprise 2-10 ligands attached covalently to a linker that links theligands in a manner that allows their multivalent binding to ligandbinding sites of Ca⁺⁺ channels. The linker spatially constrains theseinteractions to occur within dimensions defined by the linker. This andother factors increases the biologic and/or therapeutic effect of themultibinding compound as compared to the same number of ligands used inmonobinding form. The compounds of this invention are preferablyrepresented by the empirical formula (L)_(p)(X)_(q) where L, X, p and qare as defined above. This is intended to include the several ways inwhich the ligands can be linked together in order to achieve theobjective of multivalency, and a more detailed explanation is providedbelow.

[0214] As noted previously, the linker may be considered as a frameworkto which ligands are attached. Thus, it should be recognized that theligands can be attached at any suitable position on this framework, forexample, at the termini of a linear chain or at any intermediateposition thereof.

[0215] The simplest and most preferred multibinding compound is abivalent compound which can be represented as L-X-L, where L is a ligandand is the same or different and X is the linker. A trivalent compoundcould also be represented in a linear fashion, i.e., as a sequence ofrepeated units L—X—L—X—L, in which L is a ligand and is the same ordifferent at each occurrence, as is X. However, a trivalent compound canalso comprise three ligands attached to a central core, and thus berepresented as (L)₃X, where the linker X could include, for example, anaryl or cycloalkyl group. Tetravalent compounds can be represented in alinear array, L—X—L—X—L—X—L, or a branched array,

[0216] i.e., a branched construct analogous to the isomers of butane(n-butyl, iso-butyl, sec-butyl, and t-butyl). Alternatively, it could berepresented as an aryl or cycloalkyl derivative as described above withfour (4) ligands attached to the core linker.

[0217] The same considerations apply to higher multibinding compounds ofthis invention containing from 5-10 ligands. However, for multibindingagents attached to a central linker such as an aryl , cycloalkyl orheterocyclyl group, or a crown compound, there is a self-evidentconstraint that there must be sufficient attachment sites on the linkerto accommodate the number of ligands present; for example, a benzenering could not accommodate more than 6 ligands, whereas a multi-ringlinker (e.g., biphenyl) could accommodate a larger number of ligands.

[0218] The formula (L)_(p)(X)_(q) is also intended to represent a cycliccompound of formula (—L—X—), ,where n is 2-10.

[0219] All of the above variations are intended to be within the scopeof the invention defined by the formula (L)_(p)(X)_(q). Examples ofbivalent and higher-order valency compounds of this invention areprovided in FIGS. 4-20. With the foregoing in mind, a preferred linkermay be represented by the following formula:

—X′—Z—(Y′—Z)_(m)—Y″—Z—X′—

[0220] in which:

[0221] m is an integer of from 0 to 20;

[0222] X′ at each separate occurrence is —O—, —S—, —S(O)—, —S(O)₂—,—NR—, —N⁺ R R′—, —C(O)—, —C(O)O—, —C(O)NH—, —C(S), —C(S)O—, —C(S)NH— ora covalent bond, where R and R′ at each separate occurrence are asdefined below for R′and R″;

[0223] Z is at each separate occurrence selected from alkylene,substituted alkylene, alkylalkoxy, cycloalkylene, substitutedcycloalkylene, alkenylene, substituted alkenylene, alkynylene,substituted alkynylene, cycloalkenylene, substituted alkenylene,arylene, substituted arylene, heteroarylene, heterocyclene, substitutedheterocyclene, crown compounds, or a covalent bond;

[0224] Y′ and Y′ at each separate occurrence are selected from —S—S— ora covalent bond;

[0225] in which:

[0226] n is 0, 1 or 2; and

[0227] R′ and R″ at each separate occurrence are selected from hydrogen,alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, aryl, heteroaryl orheterocyclic. Additionally, the linker moiety can be optionallysubstituted at any atom therein by one or more alkyl, substituted alkyl,cycloalkyl, substituted cycloalkyl, alkenyl, substituted alkenyl,alkynyl, substituted alkynyl, aryl, heteroaryl and heterocyclic group.

[0228] As indicated above, the simplest (and preferred) construct is abivalent compound which can be represented as L—X—L, where L is a Ca⁺⁺channel ligand that is the same or different at each occurrence, and Xis the linker. Accordingly, examples of the preparation of a bivalentligand are given below as an illustration of the manner in whichmultibinding compounds of Formula I are obtained.

[0229] The reaction schemes that follow illustrate preferred linkingstrategies for linking dihydropyridine, benzothiazepine andphenylalkylamine classes of calcium channel modulators. These strategiesare intended to apply as well to any Ca⁺⁺ channel ligand that includes,or can be functionalized with groups compatible with the chosen linker(e.g., mibefradil).

[0230] As was previously discussed, the linker or linkers can beattached to different positions on the ligand molecule to achievedifferent orientations of the ligand domains and thereby facilitatemultivalency. For example, the positions that are potentially availablefor linking a dihydropyridine of formula (1), a benzothiazepine offormula (2) verapamil, a phenylalkylamine (3) and mibefradil, a tetralolderivative and selective T-channel ligand (4) are indicated by arrows inthe structures shown below.

[0231] Preferred positions of attachment suggested by known SAR areillustrated in the reaction schemes of FIGS. 4-20. The R groups shownabove are numbered to correspond to structures shown in the reactionschemes. Examples of ligands are shown in Tables 4 and 5 (Appendix).

[0232] It should be understood that a large number of Ca⁺⁺ channelligands are chiral and exhibit stereoselectivity. The most activeenantiomers are preferably used as ligands in the multibinding compoundsof this invention. The chiral resolution of enantiomers is accomplishedby well known procedures that result in the formation of diastereomericderivatives or salts, followed by conventional separation bychromatographic procedures or by fractional crystallization (see, e.g.,Bossert et al, Angew. Chem. Int. Ed. 20: 762-769 (1981) and U.S. Pat.No. 5,571,827 and references cited therein).

[0233] The ligands are covalently attached to the linker usingconventional chemical techniques. The reaction chemistries resulting insuch linkage are well known in the art and involve the coupling ofreactive functional groups present on the linker and ligand. In somecases, it may be necessary to protect portions of the ligand that arenot involved in linking reactions. Protecting groups for this purposeare well known in the art and are indicated generally in the reactionschemes by the symbols PG and PG′. Preferably, the reactive functionalgroups on the linker are selected relative to the functional groups onthe ligand that are available for coupling, or can be introduced ontothe ligand for this purpose. In some embodiments, the linker is coupledto ligand precursors, with the completion of ligand synthesis beingcarried out in a subsequent step (see, e.g., FIG. 7, Appendix). Wherefunctional groups are lacking, they can be created by suitablechemistries that are described in standard organic chemistry texts suchas J. March, “Advanced Organic Chemistry”, 4^(th) Edition(Wiley-Interscience, N.Y., 1992). Examples of the chemistry forconnecting ligands by a linker are shown in Table 3 (Appendix), where R¹and R² represent a ligand and/or the linking group. One skilled in theart will appreciate that synthetically equivalent coupling reactions canbe substituted for the reactions illustrated herein.

[0234] The linker to which the ligands or ligand precursors are attachedcomprises a “core” molecule having two or more functional groups withreactivity that is complementary to that of the functional groups on theligand. FIG. 3 illustrates the diversity of “cores” that are useful forvarying the linker size, shape, length, orientation, rigidity,acidity/basicity, hydrophobicity/hydrophilicity, hydrogen bondingcharacteristics and number of ligands connected. This pictorialrepresentation is intended only to illustrate the invention, and not tolimit its scope to the structures shown. In the Figures and reactionschemes that follow, a solid circle is used to generically represent acore molecule. The solid circle is equivalent to a linker as definedabove after reaction.

SYNTHESES OF BIVALENT COMPOUNDS

[0235] The preferred compounds of Formula I are bivalent. Accordingly,and for the purpose of simplicity, the figures and reaction schemesbelow illustrate the synthesis of bivalent Ca⁺⁺ channel modulators. Itshould be noted, however, that the same techniques can be used togenerate higher order multibinding compounds, i.e., the compounds of theinvention where p is 3-10.

[0236] Reactions performed under standard amide coupling conditions arecarried out in an inert polar solvent (e.g., DMF, DMA) in the presenceof a hindered base (e.g., TEA, DIPEA) and standard amide couplingreagents (e.g., DPPA, PyBOP, HATU, DCC).

[0237] Preparation of dihydropyridine (DHP) bivalent compounds

[0238]FIG. 4 illustrates a preferred method for linking molecules atpositions R² (R⁶) of the dihydropyridine ring. As exemplified here foramlodipine and structurally analogous molecules, a multistep HantzschDHP-cyclization is used to introduce the appropriate groups [see, e.g.,Arrowsmith et al, J. Med. Chem. 29: 1696-1702 (1986); Bossert et al,Angew. Chem. Int. Ed. Engl. 20: 762-769 (1981)).

[0239] The starting materials are a compound of formula (4) having anazide-substituted β-keto ester and a substituted benzaldehyde (5). Thesecompounds are reacted in benzene in the presence of piperidine andacetic acid to yield an intermediate product, which, when reacted with acompound of formula (6) in benzene yields DHP (7), where R² isCH₂OCH₂CH₂N₃. The ring nitrogen is protected to yield the compound offormula (8), following which the azide is reduced to an amine compound(9) using hydrogen and an appropriate catalyst, such aspalladium/calcium carbonate. Amine (9) can then be coupled to a coremolecule using a variety of well known reactions, examples of which areshown in FIG. 4 and described below. The compound of formula (9) isreacted with a bifunctional alkylating core (e.g. a dibromide linker(10)) in an inert solvent (e.g. DMF) with a hindered base (e.g. DIPEA)to produce, after deprotection, the amine linked compound of Formula I(11). Compound (11) can also be formed by reacting (9) with a dialdehydecore (12) under standard reductive amination conditions (e.g. sodiumcyanoborohydride in ethanol with acid), followed by deprotection. In yetanother type of coupling reaction, the compound of form (9) is reactedwith an activated diacid core (13) in a polar inert solvent (e.g.CH₂Cl₂) to yield, after deprotection, the amide-linked compound ofFormula I (14). The diacid may be preactivated (e.g. by using the acidchloride), or activated in situ using conventional coupling conditions(e.g. DCC, DMAP, THF). Alternatively, the compound of formula (9) can bereacted with a diisocyanate core (15) in an inert solvent (e.g. THF) toyield, after deprotection, the urea-linked compound of Formula I (16).The compound of formula (9) can also be reacted with an activatedsulfonate core (17) in an inert solvent (e.g. THF) with a hindered base(e.g. DIPEA) to yield, after deprotection, the sulfonamide linkedcompound of Formula I (18).

[0240]FIGS. 5 and 6 illustrate the preparation of dihydropyridinecompounds of Formula I having different functional linking groups atpositions R² (R⁶) of the dihydropyridine ring.

[0241] As shown in FIG. 5, a dihydropyridine with an alcohol side chainat R² is prepared by reacting a compound of formula (19) with (5) and(6) (using the same conditions as above), to make a compound of formula(20). After protection with a suitable amine-protecting group, thecompound of formula (21) can be coupled to various cores, as exemplifiedin FIG. 5. Alternatively, a compound of formula (21) can be prepared asdescribed in Alker, D and Denton, S. M., Tetrahedron, 46, 3693-3702,(1990).

[0242] The compound of formula (25) is reacted with a bifunctionalalkylating core (e.g., a diol (10)) in an inert solvent (e.g. THF) witha strong deprotonating base (e.g. NaH) to produce, after deprotection,an ether linked compound of Formula I (22). Alternatively, the compoundof formula (21) is reacted with an activated diacid core (13) in a polarinert solvent (e.g. THF) with base to yield, after deprotection, anester-linked compound of Formula I (23). Alternatively, the compound offormula (21) is reacted with a diisocyanate core (15) in an inertsolvent (e.g. THF) to yield, after deprotection, a carbamic ester-linkedcompound of Formula I (24).

[0243]FIG. 6 illustrates reactions for converting a nucleophilic sidechain into an electrophilic one. For example, the compound of formula(21) is reacted under standard chlorinating conditions (e.g. SO₂Cl₂ inthe presence of a suitable base such as imidazole in DMF) to yieldchloride (25). The compound of formula (21) is reacted with mesylchloride in an inert solvent (e.g., THF) in the presence of a base toyield mesylate (26). The compound of formula (21) is reacted withoxidating agents (e.g. CrO₃)to yield acid (27). The compound of form(21) is reacted under mild oxidating conditions (e.g. PCC/CH₂Cl₂) toyield aldehyde (28).

[0244] The electrophilic groups thus generated can then be used instandard coupling reactions such as those shown in FIGS. 4 and 5, forexample, the reaction illustrated in FIG. 6 (bottom). In this reaction,a molar equivalent of a diol core (29) is reacted with 2 molarequivalents of a mesylated dihydropyridine (26) in an inert solvent witha base to yield, after deprotection, an ether-linked compound of FormulaI (30).

[0245]FIG. 7 illustrates a preferred method for forming bivalent DHPcompounds, which involves coupling of the linker to ligand precursors,followed by synthesis of the ligand. As shown here, a molar equivalentof a diol core (29) is coupled with approximately two molar equivalentsof a chloride-substituted β diketone (31) in an inert solvent (e.g. DMF)in the presence of a strong base (e.g. NaH) to yield an ether-linkedcompound of formula (32). Compound (32) is reacted with (5) and (6), asdescribed above with reference to FIGS. 4 and 5, to form an ether-linkedcompound of Formula I (30).

[0246] Dihydropyridine dimers may be linked through an ester linkage atR⁷ or R⁸, as is shown in FIG. 8. The starting material, a t-butyl esterof formula (33) is reacted with compounds (5) and (6) as describedpreviously with reference to FIGS. 4 and 5, to yield a dihydropyridineof formula (34). Cleavage of the t-butyl group of (34) with dilute acidyields (35), which is then amine-protected to form (36). Standardactivation techniques (e.g. DCC/DMAP/THF) are used to couple the acid toa nucleophile core as previously described, for example to a diol core(29), to yield, after deprotection, an ester-linked compound of FormulaI (37).

[0247]FIG. 9 illustrates another procedure for synthesizing anester-linked dihydropyridine compound of Formula I. Here, a molarequivalent of a diol core (29) is coupled with two molar equivalents ofa diketone compound of formula (38) in an acid-catalyzedtransesterification reaction to form a compound of formula (39).Reaction of (39) with (5) and (6), as described above, yieldsester-linked (37).

[0248] Preparation of Benzothiazepine (BZT) Bivalent Compounds

[0249] Several methods for preparing bivalent benzothiazepine compoundsare illustrated in the reaction schemes shown in FIG. 10 (Appendix).According to FIG. 10, the starting material, a compound of formula (41)is prepared as described in U.S. Pat. No. 4,552,695. Compound (41) isreacted with a dihalide core (10), in an inert solvent (e.g. THF) in thepresence of a strong base (e.g. NaH) to form an amide N-linked compoundof Formula I (42). Following the linking step, the R¹² side chain can bedeprotected if necessary and acylated as described in theabove-mentioned patent.

[0250] Alternative coupling reactions are possible, including thoseshown in FIG. 4. In such instances, those skilled in the art will knowhow to modify the reaction conditions to compensate for the reducednucleophilicity of the amide nitrogen.

[0251] Alternatively, the starting material is a compound of formula(43) (where R¹² is an ether-protected hydroxyl group), which is preparedas described in the above-referenced patent. Deprotection of thiscompound yields the alcohol (44), which can be reacted with an activateddiacid core (13) in an inert solvent (e.g. DMF) to yield ester-linked(45) as shown here. Alternative coupling reactions can be used, such asthose shown in FIG. 5.

[0252] Alternatively, a compound of formula (77) may be prepared asdescribed in the above-referenced patent. Treatment of compound (77)with BBr₃ in CH₂Cl₂ affords cleavage of the aryl ether to phenol (78).Compound (78) can be selectively alkylated at the phenolic hydroxylgroup by reaction with dihalide (10) in K₂CO₃/acetone solution to yieldether-linked (79).

[0253] Preparation of Phenylalkylamine (PM) Bivalent Compounds

[0254]FIG. 11 illustrates methods for linking PM molecules (asexemplified by verapamil) (mibefradil is also considered herein to bepart of the category of phenylalkylamines). Phenyl acetonitrile (80) istreated with a basic condensing agent (e.g. sodium amide) in an inertsolvent (e.g. toluene). N-protected (81) is slowly added to the amidinesalt, as described in U.S. Pat. No. 3,261,859. After deprotection,compound (82) is coupled to an electrophilic core, for example, with adihalide core (10) in an inert solvent (e.g. DMF) in the presence of ahindered base (e.g. DIPEA) to form an amine-linked compound of Formula I(83). Other coupling reactions may be substituted for the one shownhere, e.g., those shown in FIG. 4.

[0255] Alternatively, the amidine salt of (80), is coupled toO-protected (84). After deprotection, the resulting compound (85) isreacted with a dihalide core (10), in a K₂CO₃/acetone solution to forman ether linked compound of Formula I (86). Other coupling reactionssuch as those shown in FIG. 5 may be substituted for the one shown here.

[0256] The strategies for preparing compounds of Formula I discussedabove involve coupling the ligand directly to a homobifunctional core.Another strategy that can be used with all ligands, and for thepreparation of both bivalent and higher order multibinding compounds, isto introduce a ‘spacer’ before coupling to a central core. Such a spacercan itself be selected from the same set as the possible core compounds.Examples of this linking strategy are shown in FIGS. 12 and 13, wherethe spacer is represented by a gray circle. As defined herein, thelinker comprises the spacer+core.

[0257] Referring to FIG. 12, a dihydropyridine compound of formula (21),synthesized according to FIG. 5 above, is coupled to aheterobifunctional spacer (46) having an electrophilic group (e.g., Br)and a masked nucleophilic group (e.g., protected alcohol). Theprotecting groups, PG and PG′ are different (e.g., PG is Boc and PG′ isCbz) and are capable of selective removal. The reaction is carried outin an inert solvent (e.g. DMF) in the presence of a strong base (e.g.NaH). After removal of PG′, the unmasked nucleophile (49) is coupled toan activated diacid core (13). The ring nitrogen is then deprotected toyield an ether-linked compound of Formula I (48). Core molecules withdifferent functional groups may be substituted for the one shown here(see, e.g., FIG. 5).

[0258] In another example, a dihydropyridine compound of formula (36) iscoupled to heterobifunctional spacer (49) under standard couplingconditions (e.g. DCC/DMAP/CH₂Cl₂). After removal of the spacerprotecting group, the unmasked nucleophile (50) is coupled to adialdehyde core (12) by reductive amination (or to another core withchemically compatible functional groups). After removal of theprotecting group from the ring nitrogen, an ester-linked compound offormula I (51) is obtained.

[0259]FIG. 13 illustrates the use of spacers to make bivalentbenzothiazepine compounds. As shown in FIG. 13, a compound of formula(78) is coupled to a heterobifunctional spacer (46) in basic conditions(e.g. K₂CO₃ in acetone). After removal of the spacer protecting group,the unmasked nucleophile (52) is coupled to an activated diacid core(13) using standard coupling conditions to yield an ether-linkedcompound of Formula I (53). In another example, compound (41) is coupledto heterobifunctional spacer (54) in an inert solvent solvent (e.g. DMF)in the presence of strong base (e.g. NaH). After removal of the spacerprotecting group, the resulting compound (55) is coupled to an activateddiacid core (13) to yield an N-alkylated compound of Formula I (56). Ofcourse, other suitable cores may be substituted if desired.

[0260] Compounds of Formula I where p=3-10

[0261] Compounds of Formula I of higher order valency, i.e. p>2, can beprepared by simple extension of the above strategies. As shown in FIG.14 (Appendix), compounds (58) and (61) are prepared by coupling ligandsto a central core bearing multiple functional groups. The reactionconditions are the same as described above for the preparation ofbivalent compounds, with appropriate adjustments made in the molarquantities of ligand and reagents.

[0262] Compound (36), synthesized as described above with reference toFIG. 5, is coupled to a polypeptide core with a sidechain spacer (62) tomake (63). Solid phase peptide synthesis can be used to produce a widevariety of peptidic core molecules. Techniques well known to thoseskilled in the art (including combinatorial methods) are used to varythe distance between ligand attachment sites on the core molecule, thenumber of attachment sites available for coupling, and the chemicalproperties of the core molecule. Orthogonal protecting groups are usedto selectively protect functional groups on the core molecule, thusallowing ancillary groups to be inserted into the linker of themultibinding compound and/or the preparation of “heterovalomers” (i.e.,multibinding compounds with nonidentical ligands). All of the syntheticstrategies described above employ a step in which the ligand, attachedto spacers or not, is symmetrically linked to functionally equivalentpositions on a central core. Compounds of Formula I can also besynthesized using an asymmetric linear approach. This strategy ispreferred when linking two or more ligands at different points ofconnectivity (see, e.g., FIG. 15) or when preparing heterovalomers (see,e.g., FIGS. 16 and 17).

[0263]FIG. 15 illustrates the preparation of bivalent dihydropyridinecompounds of formulae (64) and (65), wherein R² of a first ligand isattached through a linker to R⁷ of a second ligand, and the preparationof a bivalent benzothiazepine compound of formula (67), wherein thelinker is attached between R¹¹ of a first ligand and R¹³ of a secondligand. The coupling steps are carried out as described previously.

[0264] A linear strategy can also be used to prepare heterovalomers, asshown in FIGS. 16 and 17. Heterovalomers comprising different chemicalclasses of Ca⁺⁺ channel ligands (e.g., dihydropyridines andbenzothiazepines), different ligands within the same chemical class(e.g. amlodipine and isradipine) and different enantiomers of a ligand(e.g., the (+) and (−) enantiomers of a dihydropyridine) are allencompassed by the present invention.

[0265]FIG. 16 illustrates methods of preparing bivalent compoundscomprising dihydropyridine and benzothiazepine ligands in which theorientation of the ligands is varied.

[0266]FIG. 17 illustrates the preparation of a mixed agonistantagonistheterovalomer. In this example, the compound of formula (66), a Ca⁺⁺channel antagonist with an attached spacer, is coupled to the compoundof formula (75), a Ca⁺⁺ channel agonist, and deprotected to yield acompound of formula (76).

[0267] FIGS. 18-20 illustrate alternate methods of preparing bivalentcompounds comprising dihydropyridine and benzothiazepine derivatives.

[0268] Isolation and Purification of the Compounds

[0269] Isolation and purification of the compounds and intermediatesdescribed herein can be effected, if desired, by any suitable separationor purification such as, for example, filtration, extraction,crystallization, column chromatography, thin-layer chromatography,thick-layer chromatography, preparative low or high-pressure liquidchromatography or a combination of these procedures. Characterization ispreferably by NMR and mass spectroscopy.

[0270] Combinatorial Libraries

[0271] The methods described above lend themselves to combinatorialapproaches for identifying multimeric compounds which possessmultibinding properties.

[0272] Specifically, factors such as the proper juxtaposition of theindividual ligands of a multibinding compound with respect to therelevant array of binding sites on a target or targets is important inoptimizing the interaction of the multibinding compound with itstarget(s) and to maximize the biological advantage through multivalency.One approach is to identify a library of candidate multibindingcompounds with properties spanning the multibinding parameters that arerelevant for a particular target. These parameters include: (1) theidentity of ligand(s), (2) the orientation of ligands, (3) the valencyof the construct, (4) linker length, (5) linker geometry, (6) linkerphysical properties, and (7) linker chemical functional groups.Libraries of multimeric compounds potentially possessing multibindingproperties (i.e., candidate multibinding compounds) and comprising amultiplicity of such variables are prepared and these libraries are thenevaluated via conventional assays corresponding to the ligand selectedand the multibinding parameters desired. Considerations relevant to eachof these variables are set forth below:

[0273] Selection of Ligand(s):

[0274] A single ligand or set of ligands is (are) selected forincorporation into the libraries of candidate multibinding compoundswhich library is directed against a particular biological target ortargets. The only requirement for the ligands chosen is that they arecapable of interacting with the selected target(s). Thus, ligands may beknown drugs, modified forms of known drugs, substructures of known drugsor substrates of modified forms of known drugs (which are competent tointeract with the target), or other compounds. Ligands are preferablychosen based on known favorable properties that may be projected to becarried over to or amplified in multibinding forms. Favorable propertiesinclude demonstrated safety and efficacy in human patients, appropriatePK/ADME profiles, synthetic accessibility, and desirable physicalproperties such as solubility, log P, etc. However, it is crucial tonote that ligands which display an unfavorable property from among theprevious list may obtain a more favorable property through the processof multibinding compound formation; i.e., ligands should not necessarilybe excluded on such a basis. For example, a ligand that is notsufficiently potent at a particular target so as to be efficacious in ahuman patient may become highly potent and efficacious when presented inmultibinding form. A ligand that is potent and efficacious but not ofutility because of a non-mechanism-related toxic side effect may haveincreased therapeutic index (increased potency relative to toxicity) asa multibinding compound. Compounds that exhibit short in vivo half-livesmay have extended half-lives as multibinding compounds. Physicalproperties of ligands that limit their usefulness (e.g. poorbioavailability due to low solubility, hydrophobicity, hydrophilicity)may be rationally modulated in multibinding forms, providing compoundswith physical properties consistent with the desired utility.

[0275] Orientation: Selection of Ligand Attachment Points and LinkingChemistry

[0276] Several points are chosen on each ligand at which to attach theligand to the linker. The selected points on the ligand/linker forattachment are functionalized to contain complementary reactivefunctional groups. This permits probing the effects of presenting theligands to their receptor(s) in multiple relative orientations, animportant multibinding design parameter. The only requirement forchoosing attachment points is that attaching to at least one of thesepoints does not abrogate activity of the ligand. Such points forattachment can be identified by structural information when available.For example, inspection of a co-crystal structure of a proteaseinhibitor bound to its target allows one to identify one or more siteswhere linker attachment will not preclude the enzyme:inhibitorinteraction. Alternatively, evaluation of ligand/target binding bynuclear magnetic resonance will permit the identification of sitesnon-essential for ligand/target binding. See, for example, Fesik, etal., U.S. Pat. No. 5,891,643. When such structural information is notavailable, utilization of structure-activity relationships (SAR) forligands will suggest positions where substantial structural variationsare and are not allowed. In the absence of both structural and SARinformation, a library is merely selected with multiple points ofattachment to allow presentation of the ligand in multiple distinctorientations. Subsequent evaluation of this library will indicate whatpositions are suitable for attachment.

[0277] It is important to emphasize that positions of attachment that doabrogate the activity of the monomeric ligand may also be advantageouslyincluded in candidate multibinding compounds in the library providedthat such compounds bear at least one ligand attached in a manner whichdoes not abrogate intrinsic activity. This selection derives from, forexample, heterobivalent interactions within the context of a singletarget molecule. For example, consider a receptor antagonist ligandbound to its target receptor, and then consider modifying this ligand byattaching to it a second copy of the same ligand with a linker whichallows the second ligand to interact with the same receptor molecule atsites proximal to the antagonist binding site, which include elements ofthe receptor that are not part of the formal antagonist binding siteand/or elements of the matrix surrounding the receptor such as themembrane. Here, the most favorable orientation for interaction of thesecond ligand molecule with the receptor/matrix may be achieved byattaching it to the linker at a position which abrogates activity of theligand at the formal antagonist binding site. Another way to considerthis is that the SAR of individual ligands within the context of amultibinding structure is often different from the SAR of those sameligands in momomeric form.

[0278] The foregoing discussion focused on bivalent interactions ofdimeric compounds bearing two copies of the same ligand joined to asingle linker through different attachment points, one of which mayabrogate the binding/activity of the monomeric ligand. It should also beunderstood that bivalent advantage may also be attained withheterodimeric constructs bearing two different ligands that bind tocommon or different targets. For example, a 5HT₄ receptor antagonist anda bladder-selective muscarinic M₃ antagonist may be joined to a linkerthrough attachment points which do not abrogate the binding affinity ofthe monomeric ligands for their respective receptor sites. The dimericcompound may achieve enhanced affinity for both receptors due tofavorable interactions between the 5HT₄ ligand and elements of the M₃receptor proximal to the formal M₃ antagonist binding site and betweenthe M₃ ligand and elements of the 5HT₄ receptor proximal to the formal5HT₄ antagonist binding site. Thus, the dimeric compound may be morepotent and selective antagonist of overactive bladder and a superiortherapy for urinary urge incontinence.

[0279] Once the ligand attachment points have been chosen, oneidentifies the types of chemical linkages that are possible at thosepoints. The most preferred types of chemical linkages are those that arecompatible with the overall structure of the ligand (or protected formsof the ligand) readily and generally formed, stable and intrinsicallyinocuous under typical chemical and physiological conditions, andcompatible with a large number of available linkers. Amide bonds,ethers, amines, carbamates, ureas, and sulfonamides are but a fewexamples of preferred linkages.

[0280] Linkers: Spanning Relevant Multibinding Parameters ThroughSelection of Valency, Linker Length, Linker Geometry, Rigidity, PhysicalProperties. and Chemical Functional Groups

[0281] In the library of linkers employed to generate the library ofcandidate multibinding compounds, the selection of linkers employed inthis library of linkers takes into consideration the following factors:

[0282] Valency:

[0283] In most instances the library of linkers is initiated withdivalent linkers. The choice of ligands and proper juxtaposition of twoligands relative to their binding sites permits such molecules toexhibit target binding affinities and specificities more than sufficientto confer biological advantage. Furthermore, divalent linkers orconstructs are also typically of modest size such that they retain thedesirable biodistribution properties of small molecules.

[0284] Linker length:

[0285] Linkers are chosen in a range of lengths to allow the spanning ofa range of inter-ligand distances that encompass the distance preferablefor a given divalent interaction. In some instances the preferreddistance can be estimated rather precisely from high-resolutionstructural information of targets, typically enzymes and solublereceptor targets. In other instances where high-resolution structuralinformation is not available (such as 7TM G-protein coupled receptors),one can make use of simple models to estimate the maximum distancebetween binding sites either on adjacent receptors or at differentlocations on the same receptor. In situations where two binding sitesare present on the same target (or target subunit for multisubunittargets), preferred linker distances are 2-20 , with more preferredlinker distances of 3-12 . In situations where two binding sites resideon separate (e.g., protein) target sites, preferred linker distances are20-100 , with more preferred distances of 30-70.

[0286] Linker Geometry and Rigidity:

[0287] The combination of ligand attachment site, linker length, linkergeometry, and linker rigidity determine the possible ways in which theligands of candidate multibinding compounds may be displayed in threedimensions and thereby presented to their binding sites. Linker geometryand rigidity are nominally determined by chemical composition andbonding pattern, which may be controlled and are systematically variedas another spanning function in a multibinding array. For example,linker geometry is varied by attaching two ligands to the ortho, meta,and para positions of a benzene ring, or in cis-or trans-arrangements atthe 1,1- vs. 1,2- vs. 1,3- vs. 1,4-positions around a cyclohexane coreor in cis-or trans-arrangements at a point of ethylene unsaturation.Linker rigidity is varied by controlling the number and relativeenergies of different conformational states possible for the linker. Forexample, a divalent compound bearing two ligands joined by 1,8-octyllinker has many more degrees of freedom, and is therefore less rigidthan a compound in which the two ligands are attached to the 4,4′positions of a biphenyl linker.

[0288] Linker Physical Properties:

[0289] The physical properties of linkers are nominally determined bythe chemical constitution and bonding patterns of the linker, and linkerphysical properties impact the overall physical properties of thecandidate multibinding compounds in which they are included. A range oflinker compositions is typically selected to provide a range of physicalproperties (hydrophobicity, hydrophilicity, amphiphilicity,polarization, acidity, and basicity) in the candidate multibindingcompounds. The particular choice of linker physical properties is madewithin the context of the physical properties of the ligands they joinand preferably the goal is to generate molecules with favorable PK/ADMEproperties. For example, linkers can be selected to avoid those that aretoo hydrophilic or too hydrophobic to be readily absorbed and/ordistributed in vivo.

[0290] Linker Chemical Functional Groups:

[0291] Linker chemical functional groups are selected to be compatiblewith the chemistry chosen to connect linkers to the ligands and toimpart the range of physical properties sufficient to span initialexamination of this parameter.

[0292] Combinatorial Synthesis:

[0293] Having chosen a set of n ligands (n being determined by the sumof the number of different attachment points for each ligand chosen) andm linkers by the process outlined above, a library of (n!)m candidatedivalent multibinding compounds is prepared which spans the relevantmultibinding design parameters for a particular target. For example, anarray generated from two ligands, one which has two attachment points(Al, A2) and one which has three attachment points (Bl, B2, B3) joinedin all possible combinations provide for at least 15 possiblecombinations of multibinding compounds: A1-A1 A1-A2 A1-B1 A1-B2 A1-B3A2-A2 A2-B1 A2- B2 A2-B3 B1-B1 B1-B2 B1-B3 B2-B2 B2-B3 B3-B3

[0294] When each of these combinations is joined by 10 differentlinkers, a library of 150 candidate multibinding compounds results.Given the combinatorial nature of the library, common chemistries arepreferably used to join the reactive functionalies on the ligands withcomplementary reactive functionalities on the linkers. The librarytherefore lends itself to efficient parallel synthetic methods. Thecombinatorial library can employ solid phase chemistries well known inthe art wherein the ligand and/or linker is attached to a solid support.Alternatively and preferably, the combinatorial libary is prepared inthe solution phase. After synthesis, candidate multibinding compoundsare optionally purified before assaying for activity by, for example,chromatographic methods (e.g., HPLC).

[0295] Analysis of Array by Biochemical, Analytical, Pharmacological,and Computational Methods:

[0296] Various methods are used to characterize the properties andactivities of the candidate multibinding compounds in the library todetermine which compounds possess multibinding properties. Physicalconstants such as solubility under various solvent conditions andlogD/clogD values can be determined. A combination of NMR spectroscopyand computational methods is used to determine low-energy conformationsof the candidate multibinding compounds in fluid media. The ability ofthe members of the library to bind to the desired target and othertargets is determined by various standard methods, which includeradioligand displacement assays for receptor and ion channel targets,and kinetic inhibition analysis for many enzyme targets. In vitroefficacy, such as for receptor agonists and antagonists, ion channelblockers, and antimicrobial activity, can also be determined.Pharmacological data, including oral absorption, everted gutpenetration, other pharmacokinetic parameters and efficacy data can bedetermined in appropriate models. In this way, key structure-activityrelationships are obtained for multibinding design parameters which arethen used to direct future work.

[0297] The members of the library which exhibit multibinding properties,as defined herein, can be readily determined by conventional methods.First those members which exhibit multibinding properties are identifiedby conventional methods as described above including conventional assays(both in vitro and in vivo).

[0298] Second, ascertaining the structure of those compounds whichexhibit multibinding properties can be accomplished via art recognizedprocedures. For example, each member of the library can be encrypted ortagged with appropriate information allowing determination of thestructure of relevant members at a later time. See, for example, Dower,et al., International Patent Application Publication No. WO 93/06121;Brenner, et al., Proc. Natl. Acad. Sci., USA, 89:5181 (1992); Gallop, etal., U.S. Pat. No. 5,846,839; each of which are incorporated herein byreference in its entirety. Alternatively, the structure of relevantmultivalent compounds can also be determined from soluble and untaggedlibraries of candidate multivalent compounds by methods known in the artsuch as those described by Hindsgaul, et al., Canadian PatentApplication No. 2,240,325 which was published on Jul. 11, 1998. Suchmethods couple frontal affinity chromatography with mass spectroscopy todetermine both the structure and relative binding affinities ofcandidate multibinding compounds to receptors.

[0299] The process set forth above for dimeric candidate multibindingcompounds can, of course, be extended to trimeric candidate compoundsand higher analogs thereof.

[0300] Follow-Up Synthesis and Analysis of Additional Array(s):

[0301] Based on the information obtained through analysis of the initiallibrary, an optional component of the process is to ascertain one ormore promising multibinding “lead” compounds as defined by particularrelative ligand orientations, linker lengths, linker geometries, etc.Additional libraries can then be generated around these leads to providefor further information regarding structure to activity relationships.These arrays typically bear more focused variations in linker structurein an effort to further optimize target affinity and/or activity at thetarget (antagonism, partial agonism, etc.), and/or alter physicalproperties. By iterative redesign/analysis using the novel principles ofmultibinding design along with classical medicinal chemistry,biochemistry, and pharmacology approaches, one is able to prepare andidentify optimal multibinding compounds that exhibit biologicaladvantage towards their targets and as therapeutic agents.

[0302] To further elaborate upon this procedure, suitable divalentlinkers include, by way of example only, those derived from dicarboxylicacids, disulfonylhalides, dialdehydes, diketones, dihalides,diisocyanates,diamines, diols, mixtures of carboxylic acids,sulfonylhalides, aldehydes, ketones, halides, isocyanates, amines anddiols. In each case, the carboxylic acid, sulfonylhalide, aldehyde,ketone, halide, isocyanate, amine and diol functional group is reactedwith a complementary functionality on the ligand to form a covalentlinkage. Such complementary functionality is well known in the art asillustrated in the following table: COMPLEMENTARY BINDING CHEMISTRIESFirst Reactive Group Second Reactive Group Linkage hydroxyl isocyanateurethane amine epoxide -amine hydroxyamine sulfonyl halide sulfonamidecarboxyl acid amine amide hydroxyl alkyl/aryl halide ether aldehydeamine/NaCNBH₄ amine ketone amine/NaCNBH₄ amine amine isocyanate urea

[0303] The following table illustrates, by way of example, startingmaterials (identified as X-1 through X-418) that can be used to preparelinkers incorporated in the multibinding compounds of this inventionutilizing the chemistry described above. For example,1,10-decanedicarboxylic acid, X1, can be reacted with 2 equivalents of aligand carrying an amino group in the presence of a coupling reagentsuch as DCC to provide a bivalent multibinding compound of formula (I)wherein the ligands are linked via a 1,10-decanediamido linking group.

[0304] Representative ligands for use in this invention include, by wayof example, L-1 through L-21, wherein L-1=verapamil, L-2=diltiazem,L-3=benziazem, L-4=clentiazem, L-5=nicardipine, L-6=nifedipine,L-7=nilvadipine, L-8=nitredipine, L-9=nimodipine, L-10=isradipine,L-11=lacidipine, L-12=amlodipine, L-13=nisoldipine, L-14=isradipine,L-15=mibefrodil, L-16=amlodipine, L-17=felodipine, L-18=nimodipine,L-19−bepridil, L-20=SQ32,910, and L-21-SQ32,248.

[0305] Combinations of ligands (L) and linkers (X) per this inventioninclude, by way example only, homo— and hetero-dimers wherein a firstligand is selected from L-1 through L-21 above and the second ligand andlinker is selected from the following: L-1/X-1- L-1/X-2- L-1/X-3-L-1/X-4- L-1/X-5- L-1/X-6- L-1/X-7- L-1/X-8- L-1/X-9- L-1/X-10-L-1/X-11- L-1/X-12- L-1/X-13- L-1/X-14- L-1/X-15- L-1/X-16- L-1/X-17-L-1/X-18- L-1/X-19- L-1/X-20- L-1/X-21- L-1/X-22- L-1/X-23- L-1/X-24-L-1/X-25- L-1/X-26- L-1/X-27- L-1/X-28- L-1/X-29- L-1/X-30- L-1/X-31-L-1/X-32- L-1/X-33- L-1/X-34- L-1/X-35- L-1/X-36- L-1/X-37- L-1/X-38-L-1/X-39- L-1/X-40- L-1/X-41- L-1/X-42- L-1/X-43- L-1/X-44- L-1/X-45-L-1/X-46- L-1/X-47- L-1/X-48- L-1/X-49- L-1/X-50- L-1/X-51- L-1/X-52-L-1/X-53- L-1/X-54- L-1/X-55- L-1/X-56- L-1/X-57- L-1/X-58- L-1/X-59-L-1/X-60- L-1/X-61- L-1/X-62- L-1/X-63- L-1/X-64- L-1/X-65- L-1/X-66-L-1/X-67- L-1/X-68- L-1/X-69- L-1/X-70- L-1/X-71- L-1/X-72- L-1/X-73-L-1/X-74- L-1/X-75- L-1/X-76- L-1/X-77- L-1/X-78- L-1/X-79- L-1/X-80-L-1/X-81- L-1/X-82- L-1/X-83- L-1/X-84- L-1/X-85- L-1/X-86- L-1/X-87-L-1/X-88- L-1/X-89- L-1/X-90- L-1/X-91- L-1/X-92- L-1/X-93- L-1/X-94-L-1/X-95- L-1/X-96- L-1/X-97- L-1/X-98- L-1/X-99- L-1/X-100- L-1/X-101-L-1/X-102- L-1/X-103- L-1/X-104- L-1/X-105- L-1/X-106- L-1/X-107-L-1/X-108- L-1/X-109- L-1/X-110- L-1/X-111- L-1/X-112- L-1/X-113-L-1/X-114- L-1/X-115- L-1/X-116- L-1/X-117- L-1/X-118- L-1/X-119-L-1/X-120- L-1/X-121- L-1/X-122- L-1/X-123- L-1/X-124- L-1/X-125-L-1/X-126- L-1/X-127- L-1/X-128- L-1/X-129- L-1/X-130- L-1/X-131-L-1/X-132- L-1/X-133- L-1/X-134- L-1/X-135- L-1/X-136- L-1/X-137-L-1/X-138- L-1/X-139- L-1/X-140- L-1/X-141- L-1/X-142- L-1/X-143-L-1/X-144- L-1/X-145- L-1/X-146- L-1/X-147- L-1/X-148- L-1/X-149-L-1/X-150- L-1/X-151- L-1/X-152- L-1/X-153- L-1/X-154- L-1/X-155-L-1/X-156- L-1/X-157- L-1/X-158- L-1/X-159- L-1/X-160- L-1/X-161-L-1/X-162- L-1/X-163- L-1/X-164- L-1/X-165- L-1/X-166- L-1/X-167-L-1/X-168- L-1/X-169- L-1/X-170- L-1/X-171- L-1/X-172- L-1/X-173-L-1/X-174- L-1/X-175- L-1/X-176- L-1/X-177- L-1/X-178- L-1/X-179-L-1/X-180- L-1/X-181- L-1/X-182- L-1/X-183- L-1/X-184- L-1/X-185-L-1/X-186- L-1/X-187- L-1/X-188- L-1/X-189- L-1/X-190- L-1/X-191-L-1/X-192- L-1/X-193- L-1/X-194- L-1/X-195- L-1/X-196- L-1/X-197-L-1/X-198- L-1/X-199- L-1/X-200- L-1/X-201- L-1/X-202- L-1/X-203-L-1/X-204- L-1/X-205- L-1/X-206- L-1/X-207- L-1/X-208- L-1/X-209-L-1/X-210- L-1/X-211- L-1/X-212- L-1/X-213- L-1/X-214- L-1/X-215-L-1/X-216- L-1/X-217- L-1/X-218- L-1/X-219- L-1/X-220- L-1/X-221-L-1/X-222- L-1/X-223- L-1/X-224- L-1/X-225- L-1/X-226- L-1/X-227-L-1/X-228- L-1/X-229- L-1/X-230- L-1/X-231- L-1/X-232- L-1/X-233-L-1/X-234- L-1/X-235- L-1/X-236- L-1/X-237- L-1/X-238- L-1/X-239-L-1/X-240- L-1/X-241- L-1/X-242- L-1/X-243- L-1/X-244- L-1/X-245-L-1/X-246- L-1/X-247- L-1/X-248- L-1/X-249- L-1/X-250- L-1/X-251-L-1/X-252- L-1/X-253- L-1/X-254- L-1/X-255- L-1/X-256- L-1/X-257-L-1/X-258- L-1/X-259- L-1/X-260- L-1/X-261- L-1/X-262- L-1/X-263-L-1/X-264- L-1/X-265- L-1/X-266- L-1/X-267- L-1/X-268- L-1/X-269-L-1/X-270- L-1/X-271- L-1/X-272- L-1/X-273- L-1/X-274- L-1/X-275-L-1/X-276- L-1/X-277- L-1/X-278- L-1/X-279- L-1/X-280- L-1/X-281-L-1/X-282- L-1/X-283- L-1/X-284- L-1/X-285- L-1/X-286- L-1/X-287-L-1/X-288- L-1/X-289- L-1/X-290- L-1/X-291- L-1/X-292- L-1/X-293-L-1/X-294- L-1/X-295- L-1/X-296- L-1/X-297- L-1/X-298- L-1/X-299-L-1/X-300- L-1/X-301- L-1/X-302- L-1/X-303- L-1/X-304- L-1/X-305-L-1/X-306- L-1/X-307- L-1/X-308- L-1/X-309- L-1/X-310- L-1/X-311-L-1/X-312- L-1/X-313- L-1/X-314- L-1/X-315- L-1/X-316- L-1/X-317-L-1/X-318- L-1/X-319- L-1/X-320- L-1/X-321- L-1/X-322- L-1/X-323-L-1/X-324- L-1/X-325- L-1/X-326- L-1/X-327- L-1/X-328- L-1/X-329-L-1/X-330- L-1/X-331- L-1/X-332- L-1/X-333- L-1/X-334- L-1/X-335-L-1/X-336- L-1/X-337- L-1/X-338- L-1/X-339- L-1/X-340- L-1/X-341-L-1/X-342- L-1/X-343- L-1/X-344- L-1/X-345- L-1/X-346- L-1/X-347-L-1/X-348- L-1/X-349- L-1/X-350- L-1/X-351- L-1/X-352- L-1/X-353-L-1/X-354- L-1/X-355- L-1/X-356- L-1/X-357- L-1/X-358- L-1/X-359-L-1/X-360- L-1/X-361- L-1/X-362- L-1/X-363- L-1/X-364- L-1/X-365-L-1/X-366- L-1/X-367- L-1/X-368- L-1/X-369- L-1/X-370- L-1/X-371-L-1/X-372- L-1/X-373- L-1/X-374- L-1/X-375- L-1/X-376- L-1/X-377-L-1/X-378- L-1/X-379- L-1/X-380- L-1/X-381- L-1/X-382- L-1/X-383-L-1/X-384- L-1/X-385- L-1/X-386- L-1/X-387- L-1/X-388- L-1/X-389-L-1/X-390- L-1/X-391- L-1/X-392- L-1/X-393- L-1/X-394- L-1/X-395-L-1/X-396- L-1/X-397- L-1/X-398- L-1/X-399- L-1/X-400- L-1/X-401-L-1/X-402- L-1/X-403- L-1/X-404- L-1/X-405- L-1/X-406- L-1/X-407-L-1/X-408- L-1/X-409- L-1/X-410- L-1/X-411- L-1/X-412- L-1/X-413-L-1/X-414- L-1/X-415- L-1/X-416- L-1/X-417- L-1/X-418- L-2/X-1- L-2/X-2-L-2/X-3- L-2/X-4- L-2/X-5- L-2/X-6- L-2/X-7- L-2/X-8- L-2/X-9- L-2/X-10-L-2/X-11- L-2/X-12- L-2/X-13- L-2/X-14- L-2/X-15- L-2/X-16- L-2/X-17-L-2/X-18- L-2/X-19- L-2/X-20- L-2/X-21- L-2/X-22- L-2/X-23- L-2/X-24-L-2/X-25- L-2/X-26- L-2/X-27- L-2/X-28- L-2/X-29- L-2/X-30- L-2/X-31-L-2/X-32- L-2/X-33- L-2/X-34- L-2/X-35- L-2/X-36- L-2/X-37- L-2/X-38-L-2/X-39- L-2/X-40- L-2/X-41- L-2/X-42- L-2/X-43- L-2/X-44- L-2/X-45-L-2/X-46- L-2/X-47- L-2/X-48- L-2/X-49- L-2/X-50- L-2/X-51- L-2/X-52-L-2/X-53- L-2/X-54- L-2/X-55- L-2/X-56- L-2/X-57- L-2/X-58- L-2/X-59-L-2/X-60- L-2/X-61- L-2/X-62- L-2/X-63- L-2/X-64- L-2/X-65- L-2/X-66-L-2/X-67- L-2/X-68- L-2/X-69- L-2/X-70- L-2/X-71- L-2/X-72- L-2/X-73-L-2/X-74- L-2/X-75- L-2/X-76- L-2/X-77- L-2/X-78- L-2/X-79- L-2/X-80-L-2/X-81- L-2/X-82- L-2/X-83- L-2/X-84- L-2/X-85- L-2/X-86- L-2/X-87-L-2/X-88- L-2/X-89- L-2/X-90- L-2/X-91- L-2/X-92- L-2/X-93- L-2/X-94-L-2/X-95- L-2/X-96- L-2/X-97- L-2/X-98- L-2/X-99- L-2/X-100- L-2/X-101-L-2/X-102- L-2/X-103- L-2/X-104- L-2/X-105- L-2/X-106- L-2/X-107-L-2/X-108- L-2/X-109- L-2/X-110- L-2/X-111- L-2/X-112- L-2/X-113-L-2/X-114- L-2/X-115- L-2/X-116- L-2/X-117- L-2/X-118- L-2/X-119-L-2/X-120- L-2/X-121- L-2/X-122- L-2/X-123- L-2/X-124- L-2/X-125-L-2/X-126- L-2/X-127- L-2/X-128- L-2/X-129- L-2/X-130- L-2/X-131-L-2/X-132- L-2/X-133- L-2/X-134- L-2/X-135- L-2/X-136- L-2/X-137-L-2/X-138- L-2/X-139- L-2/X-140- L-2/X-141- L-2/X-142- L-2/X-143-L-2/X-144- L-2/X-145- L-2/X-146- L-2/X-147- L-2/X-148- L-2/X-149-L-2/X-150- L-2/X-151- L-2/X-152- L-2/X-153- L-2/X-154- L-2/X-155-L-2/X-156- L-2/X-157- L-2/X-158- L-2/X-159- L-2/X-160- L-2/X-161-L-2/X-162- L-2/X-163- L-2/X-164- L-2/X-165- L-2/X-166- L-2/X-167-L-2/X-168- L-2/X-169- L-2/X-170- L-2/X-171- L-2/X-172- L-2/X-173-L-2/X-174- L-2/X-175- L-2/X-176- L-2/X-177- L-2/X-178- L-2/X-179-L-2/X-180- L-2/X-181- L-2/X-182- L-2/X-183- L-2/X-184- L-2/X-185-L-2/X-186- L-2/X-187- L-2/X-188- L-2/X-189- L-2/X-190- L-2/X-191-L-2/X-192- L-2/X-193- L-2/X-194- L-2/X-195- L-2/X-196- L-2/X-197-L-2/X-198- L-2/X-199- L-2/X-200- L-2/X-201- L-2/X-202- L-2/X-203-L-2/X-204- L-2/X-205- L-2/X-206- L-2/X-207- L-2/X-208- L-2/X-209-L-2/X-210- L-2/X-211- L-2/X-212- L-2/X-213- L-2/X-214- L-2/X-215-L-2/X-216- L-2/X-217- L-2/X-218- L-2/X-219- L-2/X-220- L-2/X-221-L-2/X-222- L-2/X-223- L-2/X-224- L-2/X-225- L-2/X-226- L-2/X-227-L-2/X-228- L-2/X-229- L-2/X-230- L-2/X-231- L-2/X-232- L-2/X-233-L-2/X-234- L-2/X-235- L-2/X-236- L-2/X-237- L-2/X-238- L-2/X-239-L-2/X-240- L-2/X-241- L-2/X-242- L-2/X-243- L-2/X-244- L-2/X-245-L-2/X-246- L-2/X-247- L-2/X-248- L-2/X-249- L-2/X-250- L-2/X-251-L-2/X-252- L-2/X-253- L-2/X-254- L-2/X-255- L-2/X-256- L-2/X-257-L-2/X-258- L-2/X-259- L-2/X-260- L-2/X-261- L-2/X-262- L-2/X-263-L-2/X-264- L-2/X-265- L-2/X-266- L-2/X-267- L-2/X-268- L-2/X-269-L-2/X-270- L-2/X-271- L-2/X-272- L-2/X-273- L-2/X-274- L-2/X-275-L-2/X-276- L-2/X-277- L-2/X-278- L-2/X-279- L-2/X-280- L-2/X-281-L-2/X-282- L-2/X-283- L-2/X-284- L-2/X-285- L-2/X-286- L-2/X-287-L-2/X-288- L-2/X-289- L-2/X-290- L-2/X-291- L-2/X-292- L-2/X-293-L-2/X-294- L-2/X-295- L-2/X-296- L-2/X-297- L-2/X-298- L-2/X-299-L-2/X-300- L-2/X-301- L-2/X-302- L-2/X-303- L-2/X-304- L-2/X-305-L-2/X-306- L-2/X-307- L-2/X-308- L-2/X-309- L-2/X-310- L-2/X-311-L-2/X-312- L-2/X-313- L-2/X-314- L-2/X-315- L-2/X-316- L-2/X-317-L-2/X-318- L-2/X-319- L-2/X-320- L-2/X-321- L-2/X-322- L-2/X-323-L-2/X-324- L-2/X-325- L-2/X-326- L-2/X-327- L-2/X-328- L-2/X-329-L-2/X-330- L-2/X-331- L-2/X-332- L-2/X-333- L-2/X-334- L-2/X-335-L-2/X-336- L-2/X-337- L-2/X-338- L-2/X-339- L-2/X-340- L-2/X-341-L-2/X-342- L-2/X-343- L-2/X-344- L-2/X-345- L-2/X-346- L-2/X-347-L-2/X-348- L-2/X-349- L-2/X-350- L-2/X-351- L-2/X-352- L-2/X-353-L-2/X-354- L-2/X-355- L-2/X-356- L-2/X-357- L-2/X-358- L-2/X-359-L-2/X-360- L-2/X-361- L-2/X-362- L-2/X-363- L-2/X-364- L-2/X-365-L-2/X-366- L-2/X-367- L-2/X-368- L-2/X-369- L-2/X-370- L-2/X-371-L-2/X-372- L-2/X-373- L-2/X-374- L-2/X-375- L-2/X-376- L-2/X-377-L-2/X-378- L-2/X-379- L-2/X-380- L-2/X-381- L-2/X-382- L-2/X-383-L-2/X-384- L-2/X-385- L-2/X-386- L-2/X-387- L-2/X-388- L-2/X-389-L-2/X-390- L-2/X-391- L-2/X-392- L-2/X-393- L-2/X-394- L-2/X-395-L-2/X-396- L-2/X-397- L-2/X-398- L-2/X-399- L-2/X-400- L-2/X-401-L-2/X-402- L-2/X-403- L-2/X-404- L-2/X-405- L-2/X-406- L-2/X-407-L-2/X-408- L-2/X-409- L-2/X-410- L-2/X-411- L-2/X-412- L-2/X-413-L-2/X-414- L-2/X-415- L-2/X-416- L-2/X-417- L-2/X-418- and so on.

Utility and Testing

[0306] The multibinding compounds of this invention can be used tomodulate calcium channels in various tissues including heart, muscle,and neurons. They will typically be used for the treatment of diseasesand conditions in mammals that involve or are mediated by Ca⁺⁺ channels,such as hypertension, cardiac arrythmias, angina pectoris, cerebralischemia, congestive heart failure, migraine, Raynaud's disease, asthmaand bronchospasm, renal impairment and acute renal failure due toprolonged renal ischemia, retinal ischemia, and pain.

[0307] The multibinding compounds of this invention are tested inwell-known and reliable assays and their activities are compared withthose of the corresponding unlinked (i.e., monovalent) ligands.

[0308] Binding Affinity to Calcium Channels:

[0309] The binding affinity is determined by a radioligand competitivedisplacement assay, essentially as described in Eltze et al, Chirality2: 233-240 (1990). The ability of the present compounds to displace(+)-[³H] isradipine or a similar radioactive ligand from calcium bindingsites of guinea pig skeletal muscle T-tubule membranes is measured invitro. The binding affinity, calculated from competition curves, iscompared with that of the monovalent ligand and/or monovalentlinker-ligand conjugate.

[0310] Ca⁺⁺ Channel Activity:

[0311] The effects of compounds of this invention on channel activityare determined by measurement of whole-cell Ba⁺⁺ currents involtage-clamped Xenopus oocytes that express various types ofvoltage-gated Ca⁺⁺ channels, as described in Bezprozvanny and Tsien,Molec. Pharmacol. 48: 540-549 (1995).

[0312] Antivasoconstrictor Activity:

[0313] Antivasoconstrictor activity is determined as described inBrittain et al, Physiologist 28: 325 (1985) as the concentration of acompound required to produce 50% vasorelaxation in KCl-contracted rabbitthoracic aorta strips in the presence of calcium. Alternatively, theconcentration of a compound required to inhibit coronaryvasoconstriction induced by a thromboxane mimetic (U-46619, i.e.,9,11-methanoepoxy-PGH₂) in guinea pig Langendorff heart preparation ismeasured as described in Eltze et al, Chirality 2: 233-240 (1990).

[0314] Antihypertensive activity:

[0315] Antihypertensive activity is determined in male spontaneouslyhypertensive rats by measurement of mean arterial blood pressure(Rovnyak et al, J. Med. Chem. 35. 3254-3263 (1992)).

[0316] Tissue selectivity:

[0317] Selectivity for vascular smooth muscle as compared with cardiacmuscle can be assessed by comparing the concentration of a multibindingcompound that produces a 50% increase in coronary blood flow in anisolated guinea-pig heart with that required to inhibit myocardialcontractility. See, e.g., Osterrieder, W. and Hoick, M., J. Cardiovasc.Pharmacol. 13: 754-9 (1989; and Cremers et al, J. Cardiovasc. PharmacoL29: 692-696 (1997).

[0318] Pharmaceutical Formulations

[0319] When employed as pharmaceuticals, the compounds of Formula I areusually administered in the form of pharmaceutical compositions. Thisinvention therefore provides pharmaceutical compositions which contain,as the active ingredient, one or more of the compounds of Formula Iabove or a pharmaceutically acceptable salt thereof and one or morepharmaceutically acceptable excipients, carriers, diluents, permeationenhancers, solubilizers and adjuvants. The compounds may be administeredalone or in combination with other therapeutic agents (e.g., otherantihypertensive drugs, diuretics and the like). Such compositions areprepared in a manner well known in the pharmaceutical art (see, e.g.,Remington's Pharmaceutical Sciences, Mace Publishing Co., Philadelphia,Pa. 17^(th) Ed. (1985) and “Modem Pharmaceutics”, Marcel Dekker, Inc.3^(rd) Ed. (G. S. Banker & C. T. Rhodes, Eds.).

[0320] The compounds of Formula I may be administered by any of theaccepted modes of administration of agents having similar utilities, forexample, by oral, parenteral, rectal, buccal, intranasal or transdermalroutes. The most suitable route will depend on the nature and severityof the condition being treated. Oral administration is a preferred routefor the compounds of this invention. In making the compositions of thisinvention, the active ingredient is usually diluted by an excipient orenclosed within such a carrier which can be in the form of a capsule,sachet, paper or other container. When the excipient serves as adiluent, it can be a solid, semi-solid, or liquid material, which actsas a vehicle, carrier or medium for the active ingredient. Thus, thecompositions can be in the form of tablets, pills, powders, lozenges,sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups,aerosols (as a solid or in a liquid medium), ointments containing, forexample, up to 10% by weight of the active compound, soft and hardgelatin capsules, suppositories, sterile injectable solutions, andsterile packaged powders. Pharmaceutically acceptable salts of theactive agents may be prepared using standard procedures known to thoseskilled in the art of synthetic organic chemistry and described, e.g.,by J. March, Advanced Organic Chemistry: Reactions, Mechanisms andStructure, ₄ ^(th) Ed. (New York: Wiley-Interscience, 1992).

[0321] Some examples of suitable excipients include lactose, dextrose,sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate,alginates, tragacanth, gelatin, calcium silicate, microcrystallinecellulose, polyvinylpyrrolidone, cellulose, sterile water, syrup, andmethyl cellulose. The formulations can additionally include: lubricatingagents such as talc, magnesium stearate, and mineral oil; wettingagents; emulsifying and suspending agents; preserving agents such asmethyl- and propylhydroxy-benzoates; sweetening agents; and flavoringagents.

[0322] The compositions of the invention can be formulated so as toprovide quick, sustained or delayed release of the active ingredientafter administration to the patient by employing procedures known in theart. Controlled release drug delivery systems for oral administrationinclude osmotic pump systems and dissolutional systems containingpolymer-coated reservoirs or drug-polymer matrix formulations. Examplesof controlled release systems are given in U.S. Pat. Nos. 3,845,770;4,326,525; 4,902514; and 5,616,345. Another preferred formulation foruse in the methods of the present invention employs transdermal deliverydevices (“patches”). Such transdermal patches may be used to providecontinuous or discontinuous infusion of the compounds of the presentinvention in controlled amounts. The construction and use of transdermalpatches for the delivery of pharmaceutical agents is well known in theart. See, e.g., U.S. Pat. Nos. 5,023,252, 4,992,445 and 5,001,139. Suchpatches may be constructed for continuous, pulsatile, or on demanddelivery of pharmaceutical agents.

[0323] The compositions are preferably formulated in a unit dosage form.The term “unit dosage forms” refers to physically discrete unitssuitable as unitary dosages for human subjects and other mammals, eachunit containing a predetermined quantity of active material calculatedto produce the desired therapeutic effect, in association with asuitable pharmaceutical excipient (e.g., a tablet, capsule, ampoule).The active compound is effective over a wide dosage range and isgenerally administered in a pharmaceutically effective amount.Preferably, for oral administration, each dosage unit contains from1-250 mg of a compound of Formula I, and for parenteral administration,preferably from 0.1 to 60 mg of a compound of Formula I or apharmaceutically acceptable salt thereof. It will be understood,however, that the amount of the compound actually administered will bedetermined by a physician, in the light of the relevant circumstances,including the condition to be treated, the chosen route ofadministration, the actual compound administered and its relativeactivity, the age, weight, and response of the individual patient, theseverity of the patient's symptoms, and the like.

[0324] For preparing solid compositions such as tablets, the principalactive ingredient is mixed with a pharmaceutical excipient to form asolid preformulation composition containing a homogeneous mixture of acompound of the present invention. When referring to thesepreformulation compositions as homogeneous, it is meant that the activeingredient is dispersed evenly throughout the composition so that thecomposition may be readily subdivided into equally effective unit dosageforms such as tablets, pills and capsules.

[0325] The tablets or pills of the present invention may be coated orotherwise compounded to provide a dosage form affording the advantage ofprolonged action. For example, the tablet or pill can comprise an innerdosage and an outer dosage component, the latter being in the form of anenvelope over the former. The two components can be separated by anenteric layer which serves to resist disintegration in the stomach andpermit the inner component to pass intact into the duodenum or to bedelayed in release. A variety of materials can be used for such entericlayers or coatings, such materials including a number of polymeric acidsand mixtures of polymeric acids with such materials as shellac, cetylalcohol, and cellulose acetate.

[0326] The liquid forms in which the novel compositions of the presentinvention may be incorporated for administration orally or by injectioninclude aqueous solutions, suitably flavored syrups, aqueous or oilsuspensions, and flavored emulsions with edible oils such as corn oil,cottonseed oil, sesame oil, coconut oil, or peanut oil, as well aselixirs and similar pharmaceutical vehicles.

[0327] Compositions for inhalation or insulation include solutions andsuspensions in pharmaceutically acceptable, aqueous or organic solvents,or mixtures thereof, and powders. The liquid or solid compositions may:contain suitable pharmaceutically acceptable excipients as describedsupra. Preferably the compositions are administered by the oral or nasalrespiratory route for local or systemic effect. Compositions inpreferably pharmaceutically acceptable solvents may be nebulized by useof inert gases. Nebulized solutions may be inhaled directly from thenebulizing device or the nebulizing device may be attached to a facemask tent, or intermittent positive pressure breathing machine.Solution, suspension, or powder compositions may be administered,preferably orally or nasally, from devices which deliver the formulationin an appropriate manner.

[0328] The following formulation examples illustrate representativepharmaceutical compositions of the present invention.

FORMULATION EXAMPLE 1

[0329] Hard gelatin capsules containing the following ingredients areprepared: Quantity Ingredient (mg/capsule) Active Ingredient 30.0 Starch305.0 Magnesium stearate 5.0

[0330] The above ingredients are mixed and filled into hard gelatincapsules in 340 mg quantities.

FORMULATION EXAMPLE 2

[0331] A tablet formula is prepared using the ingredients below:Quantity Ingredient (mg/tablet) Active Ingredient 25.0 Cellulose,microcrystalline 200.0 Colloidal silicon dioxide 10.0 Stearic acid 5.0

[0332] The components are blended and compressed to form tablets, eachweighing 240 mg.

FORMULATION EXAMPLE 3

[0333] A dry powder inhaler formulation is prepared containing thefollowing components: Ingredient Weight % Active Ingredient 5 Lactose 95

[0334] The active ingredient is mixed with the lactose and the mixtureis added to a dry powder inhaling appliance.

FORMULATION EXAMPLE 4

[0335] Tablets, each containing 30 mg of active ingredient, are preparedas follows: Quantity Ingredient (mg/tablet) Active Ingredient 30.0 mgStarch 45.0 mg Microcrystalline cellulose 35.0 mg Polyvinylpyrrolidone4.0 mg (as 10% solution in sterile water) Sodium carboxymethyl starch4.5 mg Magnesium stearate 0.5 mg Talc 1.0 mg Total 120 mg

[0336] The active ingredient, starch and cellulose are passed through aNo. 20 mesh U.S. sieve and mixed thoroughly. The solution ofpolyvinylpyrrolidone is mixed with the resultant powders, which are thenpassed through a 16 mesh U.S. sieve. The granules so produced are driedat 50° C. to 60° C. and passed through a 16 mesh U.S. sieve. The sodiumcarboxymethyl starch, magnesium stearate, and talc, previously passedthrough a No. 30 mesh U.S. sieve, are then added to the granules which,after mixing, are compressed on a tablet machine to yield tablets eachweighing 120 mg.

FORMULATION EXAMPLE 5

[0337] Capsules, each containing 40 mg of medicament are made asfollows: Quantity Ingredient (mg/capsule) Active Ingredient 40.0 mgStarch 109.0 mg Magnesium stearate 1.0 mg Total 150.0 mg

[0338] The active ingredient, starch, and magnesium stearate areblended, passed through a No. 20 mesh U.S. sieve, and filled into hardgelatin capsules in 150 mg quantities.

FORMULATION EXAMPLE 6

[0339] Suppositories, each containing 25 mg of active ingredient aremade as follows: Ingredient Amount Active Ingredient 25 mg Saturatedfatty acid glycerides to 2,000 mg

[0340] The active ingredient is passed through a No. 60 mesh U.S. sieveand suspended in the saturated fatty acid glycerides previously meltedusing the minimum heat necessary. The mixture is then poured into asuppository mold of nominal 2.0 g capacity and allowed to cool.

FORMULATION EXAMPLE 7

[0341] Suspensions, each containing 50 mg of medicament per 5.0 mL doseare made as follows: Ingredient Amount Active Ingredient 50.0 mg Xanthangum 4.0 mg Sodium carboxymethyl cellulose (11%) Microcrystallinecellulose (89%) 50.0 mg Sucrose 1.75 g Sodium benzoate 10.0 mg Flavorand Color q.v. Purified water to 5.0 mL

[0342] The active ingredient, sucrose and xanthan gum are blended,passed through a No. 10 mesh U.S. sieve, and then mixed with apreviously made solution of the microcrystalline cellulose and sodiumcarboxymethyl cellulose in water. The sodium benzoate, flavor, and colorare diluted with some of the water and added with stirring. Sufficientwater is then added to produce the required volume.

FORMULATION EXAMPLE 8

[0343] Quantity Ingredient (mg/capsule) Active Ingredient 15.0 mg Starch407.0 mg Magnesium stearate 3.0 mg Total 425.0 mg

[0344] The active ingredient, starch, and magnesium stearate areblended, passed through a No. 20 mesh U.S. sieve, and filled into hardgelatin capsules in 425.0 mg quantities.

FORMULATION EXAMPLE 9

[0345] A subcutaneous formulation may be prepared as follows: IngredientQuantity Active Ingredient 5.0 mg Corn Oil 1.0 mL

[0346] Frequently, it will be desirable or necessary to introduce thepharmaceutical composition to the brain, either directly or indirectly.Direct techniques usually involve placement of a drug delivery catheterinto the host's ventricular system to bypass the blood-brain barrier.One such implantable delivery system used for the transport ofbiological factors to specific anatomical regions of the body isdescribed in U.S. Pat. No. 5,011,472 which is herein incorporated byreference.

[0347] Indirect techniques, which are generally preferred, usuallyinvolve formulating the compositions to provide for drug latentiation bythe conversion of hydrophilic drugs into lipid-soluble drugs.Latentiation is generally achieved through blocking of the hydroxy,carbonyl, sulfate, and primary amine groups present on the drug torender the drug more lipid soluble and amenable to transportation acrossthe blood-brain barrier. Alternatively, the delivery of hydrophilicdrugs may be enhanced by intra-arterial infusion of hypertonic solutionswhich can transiently open the blood-brain barrier.

SYNTHETIC EXAMPLES EXAMPLE 1

[0348] (See FIG. 4)

[0349] Preparation of a Formula I compound wherein p is 2, q is 1, andthe ligand, L, is the amlodipine moiety linked via the side chain amineto the linker, X.

[0350] Method A

[0351] Step 1

[0352] A solution of N-BOC-amlodipine [structure 9, where PG ist-butyloxycarbonyl (BOC); R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl;and R¹⁰ is H] (2 mmol), the linker molecule 1,6-dibromohexane (1 mmol),and diisopropylethylamine (0.2 mL) in DMF (3 mL) is stirred and warmedunder an inert atmosphere. The progress of the reaction is followed bytic and when reaction is complete, the solution is poured into aqueous5% NaHCO₃ and the aqueous mixture is extracted with methylene chloride.The organic extract solution is dried (Na₂SO₄), filtered andconcentrated under reduced pressure to give the crude product. Thedesired compound is obtained by purification of the crude product by useof HPLC.

[0353] Step2

[0354] A solution of the product from the preceding reaction andtrifluoroacetic acid (3 mL) in CH₂Cl₂ (5 mL) is stirred at roomtemperature. The progress of the reaction is followed by tic. Afterreaction occurs, more CH₂Cl₂ is added and the solution is washed withaqueous Na₂CO₃ and with H₂O. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound of Formula I (structure 11, where R⁶ andR⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H) is obtained bypurification of the crude product with the use of HPLC.

[0355] In similar manner, by replacing N-BOC-amlodipine in the aboveexample with other ligands of structure 9 and/or by replacing1,6-dibromohexane with other linker molecules, other compounds ofFormula I are prepared.

[0356] Method B

[0357] A solution of amlodipine [structure 9, where PG is H; R⁶ and R⁸are methyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H] (2 mmol), the linkermolecule 1,6-dibromohexane (1 mmol), and diisopropylethylamine (0.2 mL)in DMF (3 mL) is stirred and warmed under an inert atmosphere. Theprogress of the reaction is followed by tic and when reaction iscomplete, the solution is poured into aqueous 5% NaHCO₃ and the aqueousmixture is extracted with methylene chloride. The organic extractsolution is dried (Na₂SO₄), filtered and concentrated under reducedpressure to give the crude product. The desired compound of Formula I(structure 11, where R⁸ and R⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; andR¹⁰ is H) is obtained by purification of the crude product by use ofHPLC.

[0358] In similar manner, by replacing amlodipine in the above examplewith other ligands of structure 9 and/or by replacing 1,6-dibromohexanewith other linker molecules, other compounds of Formula I are prepared.

EXAMPLE 2

[0359] (see FIG. 4)

[0360] Preparation of a Formula I compound wherein p is 2, q is 1, andthe ligand, L, is the amlodipine moiety linked via the side chain amineto the linker, X, through an amide bond

[0361] Method A

[0362] Step 1

[0363] A solution of N-BOC-amlodipine [structure 9, where PG ist-butyloxycarbonyl (BOC); R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl;and R¹⁰ is H] (2 mmol), the linker molecule 3,6-dioxaoctanedioic acid (1mmol) in CH₂Cl₂ (5 mL) is prepared under argon in a flask equipped withmagnetic stirrer and a drying tube. To this solution is addeddicyclohexylcarbodiimide (solid, 2.2 mmol). The progress of the reactionis followed by tic and after reaction occurs, the reaction solution isquenched in water, aqueous sodium bicarbonate is added and the aqueousmixture is extracted with methylene chloride. The organic layer iswashed with aqueous Na₂CO₃ and with H₂O, dried (Na₂SO₄), filtered andconcentrated under reduced pressure to give the crude product. Thedesired compound is obtained by purification of the crude product withthe use of HPLC.

[0364] Step 2

[0365] A solution of the product from the preceding reaction andtrifluoroacetic acid (3 mL) in CH₂Cl₂ (5 mL) is stirred at roomtemperature. The progress of the reaction is followed by tic. Afterreaction occurs, more CH₂Cl₂ is added and the solution is washed withaqueous Na₂CO₃ and with H₂O. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound of Formula I (structure 14, where R⁶ andR⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H) is obtained bypurification of the crude product with the use of HPLC.

[0366] In similar manner, by replacing N-BOC-amlodipine in the aboveexample with other ligands of structure 9 and/or by replacing3,6-dioxaoctanedioic acid with other linker molecules, other compoundsof Formula I are prepared.

[0367] Method B

[0368] A solution of amlodipine [structure 9, where PG is H; R⁶ and R⁸are methyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H] (2 mmol), the linkermolecule 3,6-dioxaoctanedioic acid (1 mmol) in CH₂Cl₂ (5 mL) is preparedunder argon in a flask equipped with magnetic stirrer and a drying tube.To this solution is added dicyclohexylcarbodiimide (solid, 2.2 mmol).The progress of the reaction is followed by tic and after reactionoccurs, the reaction solution is quenched in water, aqueous sodiumbicarbonate is added and the aqueous mixture is extracted with methylenechloride. The organic layer is washed with aqueous Na₂CO₃ and with H₂O,dried (Na₂SO₄), filtered and concentrated under reduced pressure to givethe crude product. The desired Formula I compound (structure 14, whereR⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H) is obtainedby purification of the crude product with the use of HPLC.

[0369] In similar manner, by replacing amlodipine in the above examplewith other ligands of structure 9 and/or by replacing3,6-dioxaoctanedioic acid with other linker molecules, other compoundsof Formula I are prepared.

EXAMPLE 3

[0370] (see FIG. 4)

[0371] Preparation of a Formula I compound wherein p is 2, q is 1, andthe ligand, L, is the amlodipine moiety linked via the side chain amineto the linker, X, through a urea bond

[0372] Method A

[0373] Step 1

[0374] A solution of the linker molecule 1 ,4-diisocyanatobutane (1mmol) in CH₂Cl₂ (5 mL) containing Et₃N (0.2 mL) is stirred and cooled inan ice-water bath under an inert atmosphere. To this is added dropwise asolution of N-BOC-amlodipine [structure 9, where PG ist-butyloxycarbonyl (BOC); R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl;and R¹⁰ is H] (2 mmol) in CH₂Cl₂ (5 mL). After addition is complete, thecooling bath is removed and the reaction solution is allowed to warm toroom temperature. The progress of the reaction is followed by tic andwhen reaction has occurred, the reaction solution is quenched in cold 5%aqueous Na₂CO₃. The layers are separated and the organic layer is washedwith aqueous Na₂CO₃, with water and is dried (Na₂SO₄), filtered andconcentrated under reduced pressure to give the crude product. Thedesired compound is obtained by purification of the crude product withthe use of HPLC.

[0375] Step 2

[0376] A solution of the product from the preceding reaction andtrifluoroacetic acid (3 mL) in CH₂Cl₂ (5 mL) is stirred at roomtemperature. The progress of the reaction is followed by tic. Afterreaction occurs, more CH₂Cl₂ is added and the solution is washed withaqueous Na₂CO₃ and with H₂O. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound of Formula I (structure 16, where R⁶ andR⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H) is obtained bypurification of the crude product with the use of HPLC.

[0377] In similar manner, by replacing N-BOC-amlodipine in the aboveexample with other ligands of structure 9 and/or by replacing1,4-diisocyanatobutane with other linker molecules, other compounds ofFormula I are prepared.

[0378] Method B

[0379] A solution of the linker molecule 1,4-diisocyanatobutane (1 mmol)in CH₂Cl₂ (5 mL) containing Et₃N (0.2 mL) is stirred and cooled in anice-water bath under an inert atmosphere. To this is added dropwise asolution of amlodipine [structure 9, where PG is H; R⁶ and R⁸ aremethyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H] (2 mmol) in CH₂Cl₂ (5mL). After addition is complete, the cooling bath is removed and thereaction solution is allowed to warm to room temperature. The progressof the reaction is followed by tic and when reaction has occurred, thereaction solution is quenched in cold 5% aqueous Na₂CO₃. The layers areseparated and the organic layer is washed with aqueous Na₂CO₃, withwater and is dried (Na₂SO₄), filtered and concentrated under reducedpressure to give the crude product. The desired Formula I compound(structure 16, where R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; andR¹⁰ is H) is obtained by purification of the crude product with the useof HPLC.

[0380] In similar manner, by replacing amlodipine in the above examplewith other ligands of structure 9 and/or by replacing1,4-diisocyanatobutane with other linker molecules, other compounds ofFormula I are prepared.

EXAMPLE 4

[0381] (see FIG. 4)

[0382] Preparation of a Formula I compound wherein p is 2, q is 1, andthe ligand, L, is the amlodipine moiety linked via the side chain amineto the linker, X, through a sulfonamide bond.

[0383] Method A

[0384] Step 1

[0385] A solution of the linker molecule benzene-1,4-bis-sulfonylchloride (1 mmol) in CH₂Cl₂ (5 mL) containing Et₃N (0.2 mL) is stirredand cooled in an ice-water bath under an inert atmosphere. To this isadded dropwise a solution of N-BOC-amlodipine [structure 9, where PG ist-butyloxycarbonyl (BOC); R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl;and R¹⁰ is H] (2 mmol) in CH₂Cl₂ (5 mL). After addition is complete, thecooling bath is removed and the reaction solution is allowed to warm toroom temperature. The progress of the reaction is followed by tic andwhen reaction has occurred, the reaction solution is quenched in cold 5%aqueous Na₂CO₃. The layers are separated and the organic layer is washedwith aqueous Na₂CO₃, with water and is dried (Na₂SO₄), filtered andconcentrated under reduced pressure to give the crude product. Thedesired compound is obtained by purification of the crude product withthe use of HPLC.

[0386] Step 2

[0387] A solution of the product from the preceding reaction andtrifluoroacetic acid (3 mL) in CH₂Cl₂ (5 mL) is stirred at roomtemperature. The progress of the reaction is followed by tic. Afterreaction occurs, more CH₂Cl₂ is added and the solution is washed withaqueous Na₂CO₃ and with H₂O. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound of Formula I (structure 18, where R⁸ andR⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H) is obtained bypurification of the crude product with the use of HPLC.

[0388] In similar manner, by replacing N-BOC-amlodipine in the aboveexample with other ligands of structure 9 and/or by replacingbenzene-1,4-bissulfonyl chloride with other linker molecules, othercompounds of Formula I are prepared.

[0389] Method B

[0390] A solution of the linker molecule benzene-1,4-bissulfonylchloride (1 mmol) in CH₂Cl₂ (5 mL) containing Et₃N (0.2 mL) is stirredand cooled in an ice-water bath under an inert atmosphere. To this isadded dropwise a solution of amlodipine [structure 9, where PG is H; R⁸and R⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H] (2 mmol) inCH₂Cl₂ (5 mL). After addition is complete, the cooling bath is removedand the reaction solution is allowed to warm to room temperature. Theprogress of the reaction is followed by tic and when reaction hasoccurred, the reaction solution is quenched in cold 5% aqueous Na₂CO₃.The layers are separated and the organic layer is washed with aqueousNa₂CO₃, with water and is dried (Na₂SO₄), filtered and concentratedunder reduced pressure to give the crude product. The desired Formula Icompound (structure 18, where R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is2-Cl; and R¹⁰ is H) is obtained by purification of the crude productwith the use of HPLC.

[0391] In similar manner, by replacing amlodipine in the above examplewith other ligands of structure 9 and/or by replacingbenzene-1,4-bissulfonyl chloride with other linker molecules, othercompounds of Formula I are prepared.

EXAMPLE 5

[0392] (see FIG. 5)

[0393] Preparation of a Formula I compound wherein p is 2, q is 1, andthe ligand, L, is the 1,4-dihydropyridine moiety linked via the2-hydroxymethyl group to the linker, X, through an ether bond.

[0394] Method A

[0395] Step 1

[0396] To a mixture of sodium hydride (3.1 mmol) and dry THF (2 mL)stirred under an inert atmosphere and protected from the atmosphere witha drying tube is added a solution of the linker molecule1,4-dihydroxymethylbenzene (1 mmol) in dry THF (2 mL). To this is addeda solution of N-BOC-1,4-dihydropyridine [structure 25, where PG ist-butyloxycarbonyl (BOC); R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl;and R¹⁰ is H] (2 mmol) in dry THF (2 mL). A solution of is added and theresulting mixture is stirred at RT. The progress of the reaction isfollowed by tic and when reaction is complete, the solution is pouredinto aqueous 5% NaHCO₃ and the aqueous mixture is extracted withmethylene chloride. The organic extract solution is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound is obtained by purification of the crudeproduct by use of HPLC.

[0397] Step 2

[0398] A solution of the product from the preceding reaction andtrifluoroacetic acid (3 mL) in CH₂Cl₂ (5 mL) is stirred at roomtemperature. The progress of the reaction is followed by tic. Afterreaction occurs, more CH₂Cl₂ is added and the solution is washed withaqueous Na₂CO₃ and with H₂O. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound of Formula I (structure 22, where R⁶ andR⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H) is obtained bypurification of the crude product with the use of HPLC.

[0399] In similar manner, by replacing N-BOC-1,4-dihydropyridine in theabove example with other ligands of structure 21 and/or by replacing1,4-dihydroxymethylbenzene with other linker molecules, other compoundsof Formula I are prepared.

[0400] Method B

[0401] Step 1

[0402] To a mixture of sodium hydride (3.1 mmol) and dry THF (2 mL)stirred under an inert atmosphere and protected from the atmosphere witha drying tube is added a solution of the linker molecule1,4-dihydroxymethylbenzene (1 mmol) in dry THF (2 mL). To this is addeda solution of 1,4-dihydropyridine [structure 25, where PG is H; R⁶ andR⁸ are methyl; R⁷is ethyl; R⁹ is 2-Cl; and R¹⁰ is H] (2 mmol) in dry THF(2 mL). A solution of is added and the resulting mixture is stirred atRT. The progress of the reaction is followed by tic and when reaction iscomplete, the solution is poured into aqueous 5% NaHCO₃ and the aqueousmixture is extracted with methylene chloride. The organic extractsolution is dried (Na₂SO₄), filtered and concentrated under reducedpressure to give the crude product. The desired compound of Formula I(structure 22, where R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; andR¹⁰ is H) is obtained by purification of the crude product by use ofHPLC.

[0403] In similar manner, by replacing 1,4-dihydropyridine in the aboveexample with other ligands of structure 25 and/or by replacing1,4-dihydroxymethylbenzene with other linker molecules, other compoundsof Formula I are prepared.

EXAMPLE 6

[0404] (see FIG. 5)

[0405] Preparation of a Formula I compound wherein p is 2, q is 1, andthe ligand, L, is the 1,4-dihydropyridine moiety linked via the2-hydroxymethyl group to the linker, X, through an ester bond

[0406] Method A

[0407] Step 1

[0408] A solution of N-BOC-1,4-dihydropyridine [structure 21, where PGis t-butyloxycarbonyl (BOC); R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is2-Cl; and R¹⁰ is H] (2 mmol), the linker molecule benzene-1,4-bisaceticacid (1 mmol), and 4-dimethylaminopyridine (10 mg) in CH₂Cl₂ (5 mL) isprepared under argon in a flask equipped with magnetic stirrer and adrying tube. To this solution is added dicyclohexylcarbodiimide (solid,2.2 mmol). The progress of the reaction is followed by tic and afterreaction occurs, the reaction solution is quenched in water, aqueoussodium bicarbonate is added and the aqueous mixture is extracted withmethylene chloride. The organic layer is washed with aqueous Na₂CO₃ andwith H₂O, dried (Na₂SO₄), filtered and concentrated under reducedpressure to give the crude product. The desired compound is obtained bypurification of the crude product with the use of HPLC.

[0409] Step 2

[0410] A solution of the product from the preceding reaction andtrifluoroacetic acid (3 mL) in CH₂Cl₂ (5 mL) is stirred at roomtemperature. The progress of the reaction is followed by tic. Afterreaction occurs, more CH₂Cl₂ is added and the solution is washed withaqueous Na₂CO₃ and with H₂O. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound of Formula I (structure 23, where R⁶ andR⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H) is obtained bypurification of the crude product with the use of HPLC.

[0411] In similar manner, by replacing N-BOC-1,4-dihydropyridine in theabove example with other ligands of structure 21 and/or by replacingbenzene-1,4-bisacetic acid with other linker molecules, other compoundsof Formula I are prepared.

[0412] Method B

[0413] A solution of 1,4-dihydropyridine [structure 21, where PG is H;R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H] (2 mmol),the linker molecule benzene-1,4-bisacetic acid (1 mmol), and4-dimethylaminopyridine (10 mg) in CH₂Cl₂ (5 mL) is prepared under argonin a flask equipped with magnetic stirrer and a drying tube. To thissolution is added dicyclohexylcarbodiimide (solid, 2.2 mmol). Theprogress of the reaction is followed by tic and after reaction occurs,the reaction solution is quenched in water, aqueous sodium bicarbonateis added and the aqueous mixture is extracted with methylene chloride.The organic layer is washed with aqueous Na₂CO₃ and with H₂O, dried(Na₂SO₄), filtered and concentrated under reduced pressure to give thecrude product. The desired compound of Formula I (structure 23, where R⁶and R⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H) is obtained bypurification of the crude product with the use of HPLC.

[0414] In similar manner, by replacing 1,4-dihydropyridine in the aboveexample with other ligands of structure 21 and/or by replacingbenzene-1,4-bisacetic acid with other linker molecules, other compoundsof Formula I are prepared.

EXAMPLE 7

[0415] (see FIG. 5)

[0416] Preparation of a Formula I compound wherein p is 2, q is 1, andthe ligand, L, is the 1,4-dihydropyridine moiety linked via the2-hydroxymethyl group to the linker, X, through a carbamate bond

[0417] Method A

[0418] Step 1

[0419] A solution of the linker molecule trans-1,4-cyclohexylisocyanate(1 mmol) in CH₂Cl₂ (5 mL) containing Et₃N (0.2 mL) is stirred and cooledin an ice-water bath under an inert atmosphere. To this is addeddropwise a solution of N-BOC-1,4-dihydropyridine [structure 21, where PGis t-butyloxycarbonyl (BOC); R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is2-Cl; and R¹⁰ is H] (2 mmol) in CH₂Cl₂ (5 mL). After addition iscomplete, the cooling bath is removed and the reaction solution isallowed to warm to room temperature. The progress of the reaction isfollowed by tic and when reaction has occurred, the reaction solution isquenched in cold 5% aqueous Na₂CO₃. The layers are separated and theorganic layer is washed with aqueous Na₂CO₃, with water and is dried(Na₂SO₄), filtered and concentrated under reduced pressure to give thecrude product. The desired compound is obtained by purification of thecrude product with the use of HPLC.

[0420] Step 2

[0421] A solution of the product from the preceding reaction andtrifluoroacetic acid (3 mL) in CH₂Cl₂ (5 mL) is stirred at roomtemperature. The progress of the reaction is followed by tic. Afterreaction occurs, more CH₂Cl₂ is added and the solution is washed withaqueous Na₂CO₃ and with H₂O. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound of Formula I (structure 24, where R⁶ andR³ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H) is obtained bypurification of the crude product with the use of HPLC.

[0422] In similar manner, by replacing N-BOC-1,4-dihydropyridine in theabove example with other ligands of structure 21 and/or by replacingtrans-1,4-cyclohexylisocyanate with other linker molecules, othercompounds of Formula I are prepared.

[0423] Method B

[0424] A solution of the linker molecule trans-1,4-cyclohexylisocyanate(1 mmol) in CH₂Cl₂ (5 mL) containing Et₃N (0.2 mL) is stirred and cooledin an ice-water bath under an inert atmosphere. To this is addeddropwise a solution of 1,4-dihydropyridine [structure 21, where PG is H;R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H] (2 mmol) inCH₂Cl₂(5 mL). After addition is complete, the cooling bath is removedand the reaction solution is allowed to warm to room temperature. Theprogress of the reaction is followed by tic and when reaction hasoccurred, the reaction solution is quenched in cold 5% aqueous Na₂CO₃.The layers are separated and the organic layer is washed with aqueousNa₂CO₃, with water and is dried (Na₂SO₄), filtered and concentratedunder reduced pressure to give the crude product. The desired compoundof Formula I (structure 24, where R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹is 2-Cl; and R¹⁰ is H) is obtained by purification of the crude productwith the use of HPLC.

[0425] In similar manner, by replacing 1,4-dihydropyridine in the aboveexample with other ligands of structure 21 and/or by replacingtrans-1,4-cyclohexylisocyanate with other linker molecules, othercompounds of Formula I are prepared.

EXAMPLE 8

[0426] (see FIG. 8)

[0427] Preparation of a Formula I compound wherein p is 2, q is 1, andthe ligand, L, is the 1,4-dihydropyridine moiety linked via the3-carboxyl group to the linker, X, through an ester bond

[0428] Method A

[0429] Step 1

[0430] A solution of N-BOC-1,4-dihydropyridine (structure 36, where PGis t-butyloxycarbonyl (BOC); R², R⁶ and R⁸ are methyl; R⁹ is 2-Cl; andR¹⁰ is H] (2 mmol), the linker molecule 1,4-bis(hydroxymethyl)benzene (1mmol), and 4-dimethylaminopyridine (10 mg) in CH₂Cl₂ (5 mL) is preparedunder argon in a flask equipped with magnetic stirrer and a drying tube.To this solution is added dicyclohexylcarbodiimide (solid, 2.2 mmol).The progress of the reaction is followed by tic and after reactionoccurs, the reaction solution is quenched in water, aqueous sodiumbicarbonate is added and the aqueous mixture is extracted with methylenechloride. The organic layer is washed with aqueous Na₂CO₃ and with H₂O,dried (Na₂SO₄), filtered and concentrated under reduced pressure to givethe crude product. The desired compound is obtained by purification ofthe crude product with the use of HPLC.

[0431] Step 2

[0432] A solution of the product from the preceding reaction andtrifluoroacetic acid (3 mL) in CH₂Cl₂ (5 mL) is stirred at roomtemperature. The progress of the reaction is followed by tic. Afterreaction occurs, more CH₂Cl₂ is added and the solution is washed withaqueous Na₂CO₃ and with H₂O. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound of Formula I (structure 37, where R², R⁶and R⁸ are methyl; R⁹ is 2-Cl; and RIO is H) is obtained by purificationof the crude product with the use of HPLC.

[0433] In similar manner, by replacing N-BOC-1,4-dihydropyridine in theabove example with other ligands of structure 36 and/or by replacing1,4-bis (hydroxymethyl)benzene with other linker molecules, othercompounds of Formula I are prepared.

[0434] Method B

[0435] A solution of dihydropyridine (structure 36, where PG is H; R²,R⁶ and R⁸ are methyl; R⁹ is 2-Cl; and R¹⁰ is H] (2 mmol), the linkermolecule 1,4-bis(hydroxymethyl)benzene (1 mmol), and4-dimethylaminopyridine (10 mg) in CH₂Cl₂ (5 mL) is prepared under argonin a flask equipped with magnetic stirrer and a drying tube. To thissolution is added dicyclohexylcarbodiimide (solid, 2.2 mmol). Theprogress of the reaction is followed by tic and after reaction occurs,the reaction solution is quenched in water, aqueous sodium bicarbonateis added and the aqueous mixture is extracted with methylene chloride.The organic layer is washed with aqueous Na₂CO₃ and with H₂O, dried(Na₂SO₄), filtered and concentrated under reduced pressure to give thecrude product. The desired compound of Formula I (structure 37, whereR², R⁶ and R⁸ are methyl; R⁹ is 2-Cl; and R¹⁰ is H) is obtained bypurification of the crude product with the use of HPLC.

[0436] In similar manner, by replacing 1,4-dihydropyridine in the aboveexample with other ligands of structure 36 and/or by replacing 1,4-bis(hydroxymethyl)benzene with other linker molecules, other compounds ofFormula I are prepared.

EXAMPLE 9

[0437] (see FIG. 8)

[0438] Preparation of a Formula I compound wherein p is 2, q is 1, andthe ligand, L, is the 1,4-dihydropyridine moiety linked via the3-carboxyl group to the linker, X, through an amide bond.

[0439] Method A

[0440] Step 1

[0441] A solution of N-BOC-1,4-dihydropyridine (structure 36, where PGis t-butyloxycarbonyl (BOC); R², R⁶ and R⁸ are methyl; R⁹ is 2-Cl; andR¹⁰ is H] (2 mmol), the linker molecule 1,5-diaminopentane (1 mmol), and4-dimethylaminopyridine (10 mg) in CH₂Cl₂ (5 mL) is prepared under argonin a flask equipped with magnetic stirrer and a drying tube. To thissolution is added dicyclohexylcarbodiimide (solid, 2.2 mmol). Theprogress of the reaction is followed by tic and after reaction occurs,the reaction solution is quenched in water, aqueous sodium bicarbonateis added and the aqueous mixture is extracted with methylene chloride.The organic layer is washed with aqueous Na₂CO₃ and with H₂O, dried(Na₂SO₄), filtered and concentrated under reduced pressure to give thecrude product. The desired compound is obtained by purification of thecrude product with the use of HPLC.

[0442] Step 2

[0443] A solution of the product from the preceding reaction andtrifluoroacetic acid (3 mL) in CH₂Cl₂ (5 mL) is stirred at roomtemperature. The progress of the reaction is followed by tic. Afterreaction occurs, more CH₂Cl₂ is added and the solution is washed withaqueous Na₂CO₃ and with H₂O. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound of Formula I (structure 37a, where R², R⁶and R⁸ are methyl; R⁹ is 2-Cl; and R¹⁰ is H) is obtained by purificationof the crude product with the use of HPLC.

[0444] In similar manner, by replacing N-BOC-1,4-dihydropyridine in theabove example with other ligands of structure 36 and/or by replacing1,5-diaminopentane with other linker molecules, other compounds ofFormula I are prepared.

[0445] Method B

[0446] A solution of dihydropyridine (structure 36, where PG is H; R²,R⁶ and R⁸ are methyl; R⁹ is 2-Cl; and R¹⁰ is H] (2 mmol), the linkermolecule 1,5-diaminopentane (1 mmol), and 4-dimethylaminopyridine (10mg) in CH₂Cl₂ (5 mL) is prepared under argon in a flask equipped withmagnetic stirrer and a drying tube. To this solution is addeddicyclohexylcarbodiimide (solid, 2.2 mmol). The progress of the reactionis followed by tic and after reaction occurs, the reaction solution isquenched in water, aqueous sodium bicarbonate is added and the aqueousmixture is extracted with methylene chloride. The organic layer iswashed with aqueous Na₂CO₃ and with H₂O, dried (Na₂SO₄), filtered andconcentrated under reduced pressure to give the crude product. Thedesired Formula I compound (structure 37a, where R², R⁶ and R⁸ aremethyl; R⁹ is 2-Cl; and R¹⁰ is H) is obtained by purification of thecrude product with the use of HPLC.

[0447] In similar manner, by replacing dihydropyridine in the aboveexample with other ligands of structure 36 and/or by replacing1,5-diaminopentane with other linker molecules, other compounds ofFormula I are prepared.

EXAMPLE 10

[0448] (see FIG. 10)

[0449] Preparation of a Formula I compound wherein p is 2, q is 1, andthe ligand, L, is the benzothiazepine moiety linked via the nitrogen ofthe amide group to the linker, X

[0450] A mixture of NaH (2.1 mmol) and DMF (1 mL) is prepared under aninert atmosphere in a flask equipped with a stirring bar and a dryingtube. To this is added first a solution of benzothiazepine (structure41, where R¹² is OAc; R¹³ is Me; and R¹⁴ is H) (2 mmol) in DMF (5 mL)and then the linker molecule 1,8-dibromooctane (1 mmol). The resultingmixture is stirred and the course of the reaction is followed by thinlayer chromatography. After reaction occurs, the reaction is quenchedwith cold dilute aq. Na₂CO₃ and extracted with methylene chloride. Theorganic layer is dried (Na₂SO₄), filtered and concentrated under reducedpressure to give the crude product. The desired compound of Formula I(structure 42, where R¹² is OAc; R¹ ³ is Me; and R¹⁴ is H) is obtainedby purification of the crude product by use of HPLC.

[0451] In similar manner, by replacing the benzothiazepine in the aboveexample with other ligands of structure 41 and/or by replacing1,8-dibromooctane with other linker molecules, other compounds ofFormula I are prepared.

EXAMPLE 11

[0452] (see FIG. 10)

[0453] Preparation of a Formula I compound wherein p is 2, q is 1, andthe ligand, L, is the benzothiazepine moiety linked via the 3-hydroxylgroup to the linker, X.

[0454] Step 1

[0455] A solution of the benzothiazepine (structure 43, where PG=Ac; R¹¹is 2-(N,N-dimethylamino)ethyl; R¹³ is methyl; and R¹⁴ is H) (1 mmol) inmethanol (5 mL) is stirred with potassium carbonate. The progress of thereaction is followed by tic. After reaction occurs, the mixture isfiltered to remove solids and the filtrate is concentrated to give thecrude product. The desired compound (structure 44) is obtained bypurification of the crude product with the use of HPLC.

[0456] Step 2

[0457] A solution of the linker molecule benzene-1,4-bisacetyl chloride(2 mmol) in methylene chloride is added slowly to a solution of thebenzothiazepine [structure 44, where R¹¹ is 2-(N,N-dimethylamino)ethyl;R¹³ is methyl; and R¹⁴ is H] (2 mmols) in methylene chloride (5 mL) andpyridine (0.5 mL) in a flask equipped with a magnetic stirrer and adrying tube and which is cooled in an ice-water bath. The course of thereaction is followed by thin layer chromatography. After reactionoccurs, the reaction solution is quenched in water and the aqueousmixture is extracted with ethyl acetate. The organic layer is washedwith aqueous Na₂CO₃, with water, and is dried (Na₂SO₄), filtered andconcentrated under reduced pressure to give the crude product. Thedesired compound of Formula I [structure 45, where R“is2-(N,N-dimethylamino)ethyl; R¹³ is methyl; and R¹⁴ is H] is obtained bypurification of the crude product by use of HPLC.

[0458] In similar manner, by replacing the benzothiazepine in the aboveexample with other ligands of structure 44 and/or by replacingbenzene-1,4-bisacetyl chloride with other linker molecules, othercompounds of Formula I are prepared.

EXAMPLE 12

[0459] (see FIG. 10)

[0460] Preparation of a Formula I compound wherein p is 2, q is 1, andthe ligand, L, is the benzothiazepine moiety linked via the oxygen ofthe phenolic group to the linker, X

[0461] Step 1

[0462] A solution of the benzothiazepine [structure 77, where R¹¹ is2-(N,N-dimethylamino) ethyl; R¹² is OAc; and R¹⁴ is H] (2 mmols) inmethylene chloride (5 mL) is stirred and cooled to −78° C. under andinert atmosphere. BBr₃ (5 mmol) is added and stirring is continued asthe cooling bath is removed and the temperature of the reaction solutionis allowed to rise to room temperature. The course of the reaction isfollowed by thin layer chromatography. After reaction occurs, thereaction solution is diluted with methylene chloride and washed withcold aqueous Na₂CO₃ and with half-saturated brine. The organic layer isdried (Na₂SO₄), filtered and concentrated under reduced pressure to givethe crude product. The desired compound [structure 78, where R¹¹ is2-(N,N-dimethylamino)ethyl; R¹² is OAc; and R¹⁴ is H] is obtained bypurification of the crude product by use of HPLC.

[0463] Step 2

[0464] A solution of the benzothiazepine [structure 78, where R¹¹ is2-(N,N-dimethylamino) ethyl; R¹² is OAc; and R¹⁴ is H] (2 mmols) andlinker molecule 1,4-bisiodomethylbenzene (1 mmol) in acetone (5 mL)containing K₂CO₃ is stirred and heated at reflux temperature under aninert atmosphere. The course of the reaction is followed by thin layerchromatography. After reaction occurs, the reaction solution is dilutedwith ethyl acetate and washed with water and with aqueous Na₂CO₃. Theorganic layer is dried (Na₂SO₄), filtered and concentrated under reducedpressure to give the crude product. The desired compound of Formula I[structure 79, where R¹¹ is 2-(N,N-dimethylamino)ethyl; R¹² is OAc; andR¹⁴ is H] is obtained by purification of the crude product by use ofHPLC.

[0465] In similar manner, by replacing the benzothiazepine in the aboveexample with other ligands of structure 78 and/or by replacing1,4-bisiodomethylbenzene with other linker molecules, other compounds ofFormula I are prepared.

EXAMPLE 13

[0466] (see FIG. 11)

[0467] Preparation of a Formula I compound wherein p is 2, q is 1, andthe ligand, L, is the verapamil moiety linked via the nitrogen of theamine group to the linker, X.

[0468] A solution of N-desmethyl-verapamil (structure 82) (2 mmol), thelinker molecule 1,4-diiodobutane (1 mmol), and diisopropylethylamine(0.2 mL) in DMF (3 mL) is stirred and warmed under an inert atmosphere.The progress of the reaction is followed by tic and when reaction iscomplete, the solution is poured into aqueous 5% NaHCO₃ and the aqueousmixture is extracted with methylene chloride. The organic extractsolution is dried (Na₂SO₄), filtered and concentrated under reducedpressure to give the crude product. The desired compound of Formula I(structure 83) is obtained by purification of the crude product by useof HPLC.

EXAMPLE14

[0469] (see FIG. 11)

[0470] Preparation of a Formula I compound wherein p is 2, q is 1, andthe ligand, L, is the verapamil moiety linked via an oxygen atom to thelinker, X.

[0471] A solution of the O-desmethyl-verapamil (structure 85) (2 mmols)and linkermolecule 1,2-bis-(2-iodoethoxy)ethane (1 mmol) in acetone (5mL) containing K₂CO₃ is stirred and heated at reflux temperature underan inert atmosphere. The course of the reaction is followed by thinlayer chromatography. After reaction occurs, the reaction solution isdiluted with ethyl acetate and washed with water and with aqueousNa₂CO₃. The organic layer is dried (Na₂SO₄), filtered and concentratedunder reduced pressure to give the crude product. The desired Formula Icompound (structure 86) is obtained by purification of the crude productby use of HPLC.

EXAMPLE 15

[0472] (see FIG. 16)

[0473] Preparation of a Formula I compound wherein p is 2, q is 1, andone ligand, L,, is the 1,4-dihydropyridine moiety linked via the3-carboxyl group to the linker, X, and the second ligand, L₂, is thebenzothioazepine moiety linked to linker, X, via the hydroxyl functionof the phenolic ring.

[0474] Method A

[0475] Step 1

[0476] A solution of the benzothiazepine [structure 78, where R¹¹ is2-(N,N-dimethylamino) ethyl; R¹² is OAc; and R¹⁴ is H (see FIG. 13)] (1mmols) and linker molecule 1-iodomethyl-4-benzyloxybenzene (1 mmol) inacetone (5 mL) containing K₂CO₃ is stirred and heated at refluxtemperature under an inert atmosphere. The course of the reaction isfollowed by thin layer chromatography. After reaction occurs, thereaction solution is diluted with ethyl acetate and washed with waterand with aqueous Na₂CO₃. The organic layer is dried (Na₂SO₄), filteredand concentrated under reduced pressure to give the crude product. Thedesired compound is obtained by purification of the crude product by useof HPLC.

[0477] Step 2

[0478] Ammonium formate (96 mg, 1.5 mmol) and 10% Pd-C (50 mg) are addedto a solution of the compound obtained in the preceding reaction inmethanol (3 mL) and THF (2 mL). The mixture is stirred at roomtemperature and the progress of the reaction is monitored by tic. Afterreaction is complete, the mixture is filtered through Celite, the filterpad is rinsed with EtOAc, the combined organic layers are washedsuccessively with aq. NaHCO₃ and with half-saturated brine, thenfiltered and concentrated under reduced pressure to give the crudeproduct. The desired compound [(see FIG. 13 and 16) structure 52, whereR¹¹ is 2-(N,N-dimethylamino)ethyl; R¹² is OAc; and R¹⁴ is H] is obtainedby purification of the crude product with the use of HPLC.

[0479] Step 3

[0480] A solution of N-BOC-1,4-dihydropyridine (structure 36, where PGis t-butyloxycarbonyl (BOC); R² is 2-(N-BOC-amino)ethoxymethyl; R⁶ andR⁸ are methyl; R⁹ is 2-Cl; and R¹⁰ is H] (1 mmol), the compound[structure 52, where R¹¹ is 2-(N,N-dimethylamino)ethyl; R¹² is Oac; andR¹⁴ is H] (1 mmol) obtained in the preceding reaction (1 mmol), and4-dimethylaminopyridine (10 mg) in CH₂Cl₂ (5 mL) is prepared under argonin a flask equipped with magnetic stirrer and a drying tube. To thissolution is added dicyclohexylcarbodiimide (solid, 2.2 mmol). Theprogress of the reaction is followed by tic and after reaction occurs,the reaction solution is quenched in water, aqueous sodium bicarbonateis added and the aqueous mixture is extracted with methylene chloride.The organic layer is washed with aqueous Na₂CO₃ and with H₂O, dried(Na₂SO₄), filtered and concentrated under reduced pressure to give thecrude product. The desired compound is obtained by purification of thecrude product with the use of HPLC.

[0481] Step 4

[0482] A solution of the product from the preceding reaction andtrifluoroacetic acid (3 mL) in CH₂Cl₂ (5 mL) is stirred at roomtemperature. The progress of the reaction is followed by tic. Afterreaction occurs, more CH₂Cl₂ is added and the solution is washed withaqueous Na₂CO₃ and with H₂O. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound of Formula I [structure 68, where R² is(2-amino)ethoxymethyl; R⁶ and R⁸ are methyl; R⁹ is 2-Cl; R¹⁰ and R¹⁴ areH; R¹¹ is 2-(N,N-dimethylamino)ethyl; and R¹² is OAc] is obtained bypurification of the crude product with the use of HPLC.

[0483] In similar manner, by replacing the 1,4-dihydropyridine of theabove example with other ligands of structure 36 and/or thebenzothiazepine in the above example with other ligands of structure 52and/or by replacing 1-iodomethyl-4-benzyloxybenzene with other linkermolecules, other compounds of Formula I are prepared.

[0484] Method B

[0485] Step 1

[0486] A solution of dihydropyridine (structure 36, where PG is H; R² is2-(N-BOC-amino)ethoxymethyl; R⁶ and R⁸ are methyl; R⁹ is 2-Cl; and R¹⁰is H] (1 mmol), the compound [structure 52, where R¹¹ is2-(N,N-dimethylamino) ethyl; R¹² is OAc; and R¹⁴ is H] (1 mmol) obtainedin the preceding reaction (1 mmol), and 4-dimethylaminopyridine (10 mg)in CH₂Cl₂ (5 mL) is prepared under argon in a flask equipped withmagnetic stirrer and a drying tube. To this solution is addeddicyclohexylcarbodiimide (solid, 2.2 mmol). The progress of the reactionis followed by tic and after reaction occurs, the reaction solution isquenched in water, aqueous sodium bicarbonate is added and the aqueousmixture is extracted with methylene chloride. The organic layer iswashed with aqueous Na₂CO₃ and with H₂O, dried (Na₂SO₄), filtered andconcentrated under reduced pressure to give the crude product. Thedesired compound is obtained by purification of the crude product withthe use of HPLC.

[0487] Step 2

[0488] A solution of the product from the preceding reaction andtrifluoroacetic acid (3 mL) in CH₂Cl₂ (5 mL) is stirred at roomtemperature. The progress of the reaction is followed by tic. Afterreaction occurs, more CH₂Cl₂ is added and the solution is washed withaqueous Na₂CO₃ and with H₂O. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired Formula I compound [structure 68, where R² is(2-amino)ethoxymethyl; R⁶ and R⁸ are methyl; R⁹ is 2-Cl; R¹⁰ and R¹⁴ areH; R¹¹ is 2-(N,N-dimethylamino)ethyl; and R¹² is OAc] is obtained bypurification of the crude product with the use of HPLC.

EXAMPLE 16

[0489] (see FIG. 16)

[0490] Preparation of a Formula I compound wherein p is 2, q is 1, andone ligand, L₁, is the 1,4-dihydropyridine moiety linked via the2-hydroxymethyl group to the linker, X, and the second ligand, L₂, isthe benzothioazepine moiety linked to X via the hydroxyl function of thephenolic ring

[0491] Method A

[0492] Step 1

[0493] A mixture of NaH (1.1 mmol) and DMF (1 mL) is prepared under aninert atmosphere in a flask equipped with a stirring bar and a dryingtube. To this is added a solution of the linker molecule1-hydroxymethyl-4-benzyloxybenzene (1 mmol) in dry DMF (5 mL) and theresulting mixture is stirred for 1 hour. Then a solution of theN-BOC-1,4-dihydropyridine [(see FIG. 12) structure 21, where PG ist-butyloxycarbonyl (BOC); R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁸ is 2-Cl;and R¹⁰ is H] (1 mmol) in dry DMF (2 mL) is added. The resulting mixtureis stirred and the course of the reaction is followed by thin layerchromatography. After reaction occurs, the reaction is quenched withcold dilute aq. Na₂CO₃ and extracted with methylene chloride. Theorganic layer is dried (Na₂SO₄), filtered and concentrated under reducedpressure to give the crude product. The desired compound is obtained bypurification of the crude product by use of HPLC.

[0494] Step 2

[0495] Ammonium formate (96 mg, 1.5 mmol) and 10% Pd-C (50 mg) are addedto a solution of the compound obtained in the preceding reaction inmethanol (3 mL) and THF (2 mL). The mixture is stirred at roomtemperature and the progress of the reaction is monitored by tic. Afterreaction is complete, the mixture is filtered through Celite, the filterpad is rinsed with EtOAc, the combined organic layers are washedsuccessively with aq. NaHCO₃ and with half-saturated brine, thenfiltered and concentrated under reduced pressure to give the crudeproduct. The desired compound [(see FIG. 12 and 16) structure 47, wherePG is t-butyloxycarbonyl (BOC); R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is2-Cl; and R¹⁰ is H] is obtained by purification of the crude productwith the use of HPLC.

[0496] Step 3

[0497] Diethyl azodicarboxylate (1 mmol) is added dropwise via a syringeto a stirred solution of triphenylphosphine (1 mmol) in THF (5 mL) atroom temperature. To this is added a solution of the compound obtainedin the preceding reaction (structure 47, where PG is t-butyloxycarbonyl(BOC); R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H) (1mmol) and the benzothiazepine [structure 78, where R¹¹ is2-(N,N-dimethylamino)ethyl; R¹² is OAc; and R¹⁴ is H] (1 mmol) in THF (3mL). The resulting solution is stirred at RT and the progress of thereaction is followed by tic. After reaction occurs, solvent is removedby evaporation under reduced pressure and the residue is purified byHPLC, giving the desired compound.

[0498] Step 4

[0499] A solution of the product from the preceding reaction andtrifluoroacetic acid (3 mL) in CH₂Cl₂ (5 mL) is stirred at roomtemperature. The progress of the reaction is followed by tic. Afterreaction occurs, more CH₂Cl₂ is added and the solution is washed withaqueous Na₂CO₃ and with H₂O. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound of Formula I [structure 69, where R⁶ andR⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; R¹⁰ and R¹⁴ are H; R¹¹ is2-(N,N-dimethylamino)ethyl; and R¹² is OAc] is obtained by purificationof the crude product with the use of HPLC.

[0500] In similar manner, by replacing the 1,4-dihydropyridine of theabove example with other ligands of structure 47 and/or thebenzothiazepine in the above example with other ligands of structure 78and/or by replacing 1-bromomethyl-4-benzyloxybenzene with other linkermolecules, other compounds of Formula I are prepared.

[0501] Method B

[0502] Step 1

[0503] A mixture of NaH (2.1 mmol) and DMF (1 mL) is prepared under aninert atmosphere in a flask equipped with a stirring bar and a dryingtube. To this is added a solution of the linker molecule1-hydroxymethyl-4-benzyloxybenzene (1 mmol) in dry DMF (5 mL) and theresulting mixture is stirred for 1 hour. Then a solution of the1,4-dihydropyridine [(see FIG. 12) structure 21, where PG is H; R⁶ andR⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; and RI” is H] (1 mmol) in dryDMF (2 mL) is added. The resulting mixture is stirred and the course ofthe reaction is followed by thin layer chromatography. After reactionoccurs, the reaction is quenched with cold dilute aq. Na₂CO₃ andextracted with methylene chloride. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound is obtained by purification of the crudeproduct by use of HPLC.

[0504] Step 2

[0505] Ammonium formate (96 mg, 1.5 mmol) and 10% Pd-C (50 mg) are addedto a solution of the compound obtained in the preceding reaction inmethanol (3 mL) and THF (2 mL). The mixture is stirred at roomtemperature and the progress of the reaction is monitored by tic. Afterreaction is complete, the mixture is filtered through Celite, the filterpad is rinsed with EtOAc, the combined organic layers are washedsuccessively with aq. NaHCO₃ and with half-saturated brine, thenfiltered and concentrated under reduced pressure to give the crudeproduct. The desired compound [(see FIG. 12 and 16) structure 47, wherePG is H; R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H] isobtained by purification of the crude product with the use of HPLC.

[0506] Step 3

[0507] Diethyl azodicarboxylate (1 mmol) is added dropwise via a syringeto a stirred solution of triphenylphosphine (1 mmol) in THF (5 mL) atroom temperature. To this is added a solution of the compound obtainedin the preceding reaction (structure 47, where PG is H; R⁶ and R⁸ aremethyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H) (1 mmol) and thebenzothiazepine [structure 78, where R¹¹ is 2-(N,N-dimethylamino)ethyl;R¹² is OAc; and R¹⁴ is H] (1 mmol) in THF (3 mL). The resulting solutionis stirred at RT and the progress of the reaction is followed by tic.After reaction occurs, solvent is removed by evaporation under reducedpressure and the residue is purified by HPLC, giving the desiredcompound of Formula I [structure 69, where R⁶ and R⁸ are methyl; R⁷ isethyl; R⁹ is 2-Cl; R¹⁰ and R¹⁴ are H; R¹⁰ is 2-(N,N-dimethylamino)ethyl; and R¹² is OAc].

EXAMPLE 17

[0508] (see FIG. 16)

[0509] Preparation of a Formula I compound wherein p is 2, q is 1, andone ligand, L,, is the 1,4-dihydropyridine moiety linked via a2-aminomethyl group to the linker, X, and the second ligand, L₂, is thebenzothioazepine moiety linked to X via the amide nitrogen of thethioazepine ring.

[0510] Method A

[0511] Step 1

[0512] A mixture of NaH (1.1 mmol) and DMF (1 mL) is prepared under aninert atmosphere in a flask equipped with a stirring bar and a dryingtube. To this is first added a solution of the benzothiazepine (see FIG.13 structure 41, where R¹² is OAc; R¹³ is methyl; and R¹⁴ is H) (1 mmol)(1 mmol) in dry DMF (3 mL) followed by the linker molecule1-bromomethyl-4-(N-Cbz-N-methyl) aminobenzene (1 mmol) in dry DMF (1mL). The resulting mixture is stirred and the course of the reaction isfollowed by thin layer chromatography. After reaction occurs, thereaction is quenched with cold dilute aq. Na₂CO₃ and extracted withmethylene chloride. The organic layer is dried (Na₂SO₄), filtered andconcentrated under reduced pressure to give the crude product. Thedesired compound is obtained by purification of the crude product by useof HPLC.

[0513] Step 2

[0514] Ammonium formate (96 mg, 1.5 mmol) and 10% Pd-C (50 mg) are addedto a solution of the compound obtained in the preceding reaction inmethanol (3 mL) and THF (2 mL). The mixture is stirred at roomtemperature and the progress of the reaction is monitored by tic. Afterreaction is complete, the mixture is filtered through Celite, the filterpad is rinsed with EtOAc, the combined organic layers are washedsuccessively with aq. NaHCO₃ and with half-saturated brine, thenfiltered and concentrated under reduced pressure to give the crudeproduct. The desired compound [(see FIG. 13 and 16) structure 55, whereR¹² is OAc; R¹³ is methyl; and R¹⁴ is H] is obtained by purification ofthe crude product with the use of HPLC.

[0515] Step 3

[0516] A solution of the compound (structure 55, where R¹² is OAc; R¹³is methyl; and R¹⁴ is H) from the preceding reaction (1 mmol) and theN-BOC-1,4-dihydropyridine [structure 25 (Alker, D.; Swanson, A. G.Tetrahedron Lett. 1990, 31, 1479-1482), where PG is t-butyloxycarbonyl(BOC); R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H] (1mmol) and diisopropylethylamine (0.2 mL) in DMF (3 mL) is stirred andwarmed under an inert atmosphere. The progress of the reaction isfollowed by tic and when reaction is complete, the solution is pouredinto aqueous 5% NaHCO₃ and the aqueous mixture is extracted withmethylene chloride. The organic extract solution is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound is obtained by purification of the crudeproduct by use of HPLC.

[0517] Step 4

[0518] A solution of the product from the preceding reaction andtrifluoroacetic acid (3 mL) in CH₂Cl₂ (5 mL) is stirred at roomtemperature. The progress of the reaction is followed by tic. Afterreaction occurs, more CH₂Cl₂ is added and the solution is washed withaqueous Na₂CO₃ and with H₂O. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound of Formula I [structure 70, where R⁶ andR⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; R¹⁰ and R¹⁴ are H; R¹² is Oac;and R¹³ is methyl] is obtained by purification of the crude product withthe use of HPLC.

[0519] In similar manner, by replacing the 1,4-dihydropyridine of theabove example with other ligands of structure 25 and/or thebenzothiazepine in the above example with other ligands of structure 55and/or by replacing 1-bromomethyl-4-( N-Cbz-N-methyl)aminobenzene [instep (1) of this example] with other linker molecules, other compoundsof Formula I are prepared.

[0520] Method B

[0521] A solution of the compound (structure 55, where R¹² is OAc; R¹³is methyl; and R¹⁴ is H) from the preceding reaction (1 mmol) and the1,4-dihydropyridine [structure 25 (Alker, D.; Swanson, A. G. TetrahedronLett. 1990, 31, 1479-1482), where PG is H; R⁶ and R⁸ are methyl; R⁷ isethyl; R⁹ is 2-Cl; and R¹⁰ is H] (1 mmol) and diisopropylethylamine (0.2mL) in DMF (3 mL) is stirred and warmed under an inert atmosphere. Theprogress of the reaction is followed by tic and when reaction iscomplete, the solution is poured into aqueous 5% NaHCO₃ and the aqueousmixture is extracted with methylene chloride. The organic extractsolution is dried (Na₂SO₄), filtered and concentrated under reducedpressure to give the crude product. The desired Formula I compound[structure 70, where R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; R¹⁰and R¹⁴ are H; R¹² is Oac; and R¹³ is methyl] is obtained bypurification of the crude product by use of HPLC.

EXAMPLE 18

[0522] (see FIG. 16)

[0523] Preparation of a Formula I compound wherein p is 2, q is 1, andone ligand, L₁, is the 1,4-dihydropyridine moiety linked via a2-hydroxymethyl group to the linker, X, and the second ligand, L₂, isthe benzothioazepine moiety linked to X via the amide nitrogen of thethioazepine ring

[0524] Method A

[0525] Step 1

[0526] A solution, cooled to the temperature of an ice-water bath,containing the N-BOC-1,4-dihydropyridine {structure 47 [see example 16,method A, step 2)] where PG is BOC; R⁶ and R⁸ are methyl; R⁷ is ethyl;R⁹ is 2-Cl; and R¹⁰ is H} (1 mmol), triphenylphosphine (1.5 mmol), andcarbon tetrabromide (2 mmol) in CH₂Cl₂ (10 mL) is prepared and isstirred. The cooling bath is removed and the solution is stirred at roomtemperature. The progress of the reaction is followed by tic and afterreaction occurs, the solution is diluted with additional CH₂Cl₂, washedwith aqueous 5% NaHCO₃, with water and with half-saturated brine. Theorganic layer is separated, dried (Na₂SO₄), filtered and concentratedunder reduced pressure to give the crude product. The desired compound(structure 59 where PG is t-butyloxycarbonyl (BOC); R⁶ and R⁸ aremethyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H) is obtained bypurification of the crude product by use of HPLC.

[0527] Step 2

[0528] A mixture of NaH (1.1 mmol) and DMF (1 mL) is prepared under aninert atmosphere in a flask equipped with a stirring bar and a dryingtube. To this is added a solution of the N-BOC-1,4-dihydropyridine[structure 59, where PG is t-butyloxycarbonyl (BOC); R⁶ and R⁸ aremethyl; R⁷ is ethyl; R⁹ is 2-Cl; and R¹⁰ is H] (1 mmol) in DMF (3 mL)followed by a solution of the benzothiazepine (structure 41, where R¹²is OAc; R¹³ is methyl; and R¹⁴ is H) (1 mmol) in dry DMF (3 mL). Theresulting mixture is stirred and the course of the reaction is followedby thin layer chromatography. After reaction occurs, the reaction isquenched with cold dilute aq. Na₂CO₃ and extracted with methylenechloride. The organic layer is dried (Na₂SO₄), filtered and concentratedunder reduced pressure to give the crude product. The desired compoundis obtained by purification of the crude product by use of HPLC.

[0529] Step 3

[0530] A solution of the product from the preceding reaction andtrifluoroacetic acid (3 mL) in CH₂Cl₂ (5 mL) is stirred at roomtemperature. The progress of the reaction is followed by tic. Afterreaction occurs, more CH₂Cl₂ is added and the solution is washed withaqueous Na₂CO₃ and with H₂O. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound of Formula I [structure 71, where R⁶, R⁸and R¹³ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; R¹⁰ and R¹⁴ are H; R¹² isOAc] is obtained by purification of the crude product with the use ofHPLC.

[0531] In similar manner, by replacing the 1,4-dihydropyridine of theabove example with other ligands of structure 59 and/or thebenzothiazepine in the above example with other ligands of structure 41and/or by replacing 1-bromomethyl-4-benzyloxybenzene [in example 16]with other linker molecules, other compounds of Formula I are prepared.

[0532] Method B

[0533] Step 1

[0534] A solution, cooled to the temperature of an ice-water bath,containing the 1,4-dihydropyridine {structure 47 [see example 16, methodB, step 2] where PG is H; R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl;and R¹⁰ is H} (1 mmol), triphenylphosphine (1.5 mmol), and carbontetrabromide (2 mmol) in CH₂CO₂ (10 mL) is prepared and is stirred. Thecooling bath is removed and the solution is stirred at room temperature.The progress of the reaction is followed by tic and after reactionoccurs, the solution is diluted with additional CH₂Cl₂, washed withaqueous 5% NaHCO₃, with water and with half-saturated brine. The organiclayer is separated, dried (Na₂SO₄), filtered and concentrated underreduced pressure to give the crude product. The desired compound(structure 59 where PG is H; R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is2-Cl; and R¹⁰ is H) is obtained by purification of the crude product byuse of HPLC.

[0535] Step 2

[0536] A mixture of NaH (1.1 mmol) and DMF (1mL) is prepared under aninert atmosphere in a flask equipped with a stirring bar and a dryingtube. To this is added a solution of the 1,4-dihydropyridine [structure59, where PG is H; R⁶ and R⁸ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; andR¹⁰ is H] (1 mmol) in DMF (3 mL) followed by a solution of thebenzothiazepine (structure 41, where R¹² is OAc; R¹³ is methyl; and R¹⁴is H) (1 mmol) in dry DMF (3 mL). The resulting mixture is stirred andthe course of the reaction is followed by thin layer chromatography.After reaction occurs, the reaction is quenched with cold dilute aq.Na₂CO₃ and extracted with methylene chloride. The organic layer is dried(Na₂SO₄), filtered and concentrated under reduced pressure to give thecrude product. The desired compound of Formula I [structure 71, whereR⁶, R³ and R¹³ are methyl; R⁷ is ethyl; R⁹ is 2-Cl; R¹⁰ and R¹⁴ are H;R¹² is OAc] is obtained by purification of the crude product by use ofHPLC.

[0537] In similar manner, by replacing the 1,4-dihydropyridine of theabove example with other ligands of structure 59 and/or thebenzothiazepine in the above example with other ligands of structure 41and/or by replacing 1-bromomethyl-4-benzyloxybenzene [in example 16,Method A, Part A(1.)] with other linker molecules, other compounds ofFormula I are prepared.

EXAMPLE 19

[0538] (see FIG. 16)

[0539] Preparation of a Formula I compound wherein p is 2, q is 1, andone ligand, L,, is the 1,4-dihydropyridine moiety linked via the3-carboxyl group to the linker, X, and the second ligand, L₂, is thebenzothioazepine moiety linked toX via the amide nitrogen of thethioazepine ring.

[0540] Method A

[0541] Step 1

[0542] A solution of N-BOC-1,4-dihydropyridine (structure 36, where PGis t-butyloxycarbonyl (BOC); R², R⁶ and R⁸ are methyl; R⁹ is 2-Cl; andR¹⁰ is H] (1 mmol), the linker molecule1-hydroxymethyl-4-bromomethylbenzene (1 mmol), and4-dimethylaminopyridine (10 mg) in CH₂Cl₂ (5 mL) is prepared under argonin a flask equipped with magnetic stirrer and a drying tube. To thissolution is added dicyclohexylcarbodiimide (solid, 2.2 mmol). Theprogress of the reaction is followed by tic and after reaction occurs,the reaction solution is quenched in water, aqueous sodium bicarbonateis added and the aqueous mixture is extracted with methylene chloride.The organic layer is washed with aqueous Na₂CO₃ and with H₂O, dried(Na₂SO₄), filtered and concentrated under reduced pressure to give thecrude product. The desired compound (structure 72, where PG ist-butyloxycarbonyl (BOC); R², R⁶ and R³ are methyl; R⁹ is 2-Cl; and R¹⁰is H) is obtained by purification of the crude product with the use ofHPLC.

[0543] Step 2

[0544] A mixture of NaH (1.1 mmol) and DMF (1 mL) is prepared under aninert atmosphere in a flask equipped with a stirring bar and a dryingtube. To this is added a solution of the benzothiazepine (structure 41,where R¹² is OAc; R¹³ is methyl; and R¹⁴ is H) (1 mmol) in dry DMF (3mL) followed by a solution of the N-BOC-1,4-dihydropyridine [structure72, where PG is t-butyloxycarbonyl (BOC); R², R⁶ and R⁸ are methyl; R⁹is 2-Cl; and R¹⁰ is H] (1 mmol) in DMF (3 mL). The resulting mixture isstirred and the course of the reaction is followed by thin layerchromatography. After reaction occurs, the reaction is quenched withcold dilute aq. Na₂CO₃ and extracted with methylene chloride. Theorganic layer is dried (Na₂SO₄), filtered and concentrated under reducedpressure to give the crude product. The desired compound is obtained bypurification of the crude product by use of

[0545] Step 3

[0546] A solution of the product from the preceding reaction andtrifluoroacetic acid (3 mL) in CH₂Cl₂ (5 mL) is stirred at roomtemperature. The progress of the reaction is followed by tIc. Afterreaction occurs, more CH₂Cl₂ is added and the solution is washed withaqueous Na₂CO₃ and with H₂O. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound of Formula I [structure 73, where R², R⁶,R⁸ and R¹³ are methyl; R⁹ is 2-Cl; R¹⁰ and R¹⁴ are H; and R¹² is OAc] isobtained by purification of the crude product with the use of HPLC.

[0547] In similar manner, by replacing the 1,4-dihydropyridine of theabove example with other ligands of structure 72 and/or thebenzothiazepine in the above example with other ligands of structure 41and/or by replacing 1-hydroxymethyl-4-bromomethylbenzene with otherlinker molecules, other compounds of Formula I are prepared.

[0548] Method B

[0549] Step 1

[0550] A solution of 1,4-dihydropyridine (structure 36, where PG is H;R², R⁶ and R⁸ are methyl; R⁹ is 2-Cl; and R¹⁰ is H] (1 mmol), the linkermolecule 1-hydroxymethyl-4-bromomethylbenzene (1 mmol), and4-dimethylaminopyridine (10 mg) in CH₂Cl₂ (5 mL) is prepared under argonin a flask equipped with magnetic stirrer and a drying tube. To thissolution is added dicyclohexylcarbodiimide (solid, 2.2 mmol). Theprogress of the reaction is followed by tic and after reaction occurs,the reaction solution is quenched in water, aqueous sodium bicarbonateis added and the aqueous mixture is extracted with methylene chloride.The organic layer is washed with aqueous Na₂CO₃ and with H₂O, dried(Na₂SO₄), filtered and concentrated under reduced pressure to give thecrude product. The desired compound (structure 72, where PG is H; R², R⁶and R⁸ are methyl; R⁹ is 2-Cl; and R¹⁰ is H) is obtained by purificationof the crude product with the use of HPLC.

[0551] Step 2

[0552] A mixture of NaH (1.1 mmol) and DMF (1 mL) is prepared under aninert atmosphere in a flask equipped with a stirring bar and a dryingtube. To this is added a solution of the benzothiazepine (structure 41,where R¹² is OAc; R¹³ is methyl; and R¹⁴ is H) (1 mmol) in dry DMF (3mL) followed by a solution of the dihydropyridine [structure 72, wherePG is H; R², R⁶ and R⁸ are methyl; R⁹ is 2-Cl; and R¹⁰ is H] (1 mmol) inDMF (3 mL). The resulting mixture is stirred and the course of thereaction is followed by thin layer chromatography. After reactionoccurs, the reaction is quenched with cold dilute aq. Na₂CO₃ andextracted with methylene chloride. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired Formula I compound [structure 73, where R², R⁶, R⁸and R¹³ are methyl; R⁹ is 2-Cl; R¹⁰ and R¹⁴ are H; and R¹² is OAc] isobtained by purification of the crude product by use of HPLC.

[0553] In similar manner, by replacing the 1,4-dihydropyridine of theabove example with other ligands of structure 72 and/or thebenzothiazepine in the above example with other ligands of structure 41and/or by replacing 1-hydroxymethyl-4-bromomethylbenzene with otherlinker molecules, other compounds of Formula I are prepared.

EXAMPLE 20

[0554] (see FIG. 17)

[0555] Preparation of a Formula I compound wherein p is 2, q is 1, andone ligand, L₁, is the 1,4-dihydropyridine moiety linked via a 6-aminogroup to the linker, X, and the second ligand, L₂, is thebenzthioazepine moiety linked to X via the hydroxyl function of thephenolic ring.

[0556] Method A

[0557] Step 1

[0558] A solution, cooled to the temperature of an ice-water bath,containing the benzothioazepine {structure 52 [see example 9], where R¹¹is 2-(N,N-dimethylamino) ethyl; R¹² is OAc; and R¹⁴ is H} (1 mmol),triphenylphosphine (1.5 mmol), and carbon tetrabromide (2 mmol) inCH₂Cl₂ (10 mL) is prepared and is stirred. The cooling bath is removedand the solution is stirred at room temperature. The progress of thereaction is followed by tic and after reaction occurs, the solution isdiluted with additional CH₂Cl₂, washed with aqueous 5% NaHCO₃, withwater and with half-saturated brine. The organic layer is separated,dried (Na₂SO₄), filtered and concentrated under reduced pressure to givethe crude product. The desired compound [structure 66 where R¹′ is2-(N,N-dimethylamino)ethyl; R¹² is OAc; and R¹⁴ is H] is obtained bypurification of the crude product by use of HPLC.

[0559] Step 2

[0560] A solution of the compound [structure 66 where R¹¹ is2-(N,N-dimethylamino)-ethyl; R¹² is OAc; and R¹⁴ is H] (1 mmol) preparedin the preceding reaction, the N-BOC-1,4-dihydropyridine [structure 75,where PG is t-butyloxycarbonyl (BOC)] (1 mmol), anddiisopropylethylamine (0.2 mL) in DMF (5 mL) is stirred and warmed underan inert atmosphere. The progress of the reaction is followed by tic andwhen reaction is complete, the solution is poured into aqueous 5% NaHCO₃and the aqueous mixture is extracted with methylene chloride. Theorganic extract solution is dried (Na₂SO₄), filtered and concentratedunder reduced pressure to give the crude product. The desired compoundis obtained by purification of the crude product by use of HPLC.

[0561] Step 3

[0562] A solution of the product from the preceding reaction andtrifluoroacetic acid (3 mL) in CH₂CO₂ (5 mL) is stirred at roomtemperature. The progress of the reaction is followed by tic. Afterreaction occurs, more CH₂Cl₂ is added and the solution is washed withaqueous Na₂CO₃ and with H₂O. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound of Formula I [structure 76, where R¹¹ is2-(N,N-dimethylamino)ethyl; R¹² is OAc; and R¹⁴ is H] is obtained bypurification of the crude product with the use of HPLC.

[0563] In similar manner, by replacing the benzothiazepine in the aboveexample with other ligands of structure 66 and/or by replacing thelinker molecule used to prepare 52 with other linker molecules, othercompounds of Formula I are prepared.

[0564] Method B

[0565] Step 1

[0566] A solution, cooled to the temperature of an ice-water bath,containing the benzothioazepine {structure 52 [see example 9, method A,step 2], where R¹¹ is 2-(N,N-dimethylamino)ethyl; R¹² is OAc; and R¹⁴ isH} (1 mmol), triphenylphosphine (1.5 mmol), and carbon tetrabromide (2mmol) in CH₂Cl₂ (10 mL) is prepared and is stirred. The cooling bath isremoved and the solution is stirred at room temperature. The progress ofthe reaction is followed by tic and after reaction occurs, the solutionis diluted with additional CH₂Cl₂, washed with aqueous 5% NaHCO₃, withwater and with half-saturated brine. The organic layer is separated,dried (Na₂SO₄), filtered and concentrated under reduced pressure to givethe crude product. The desired compound [structure 66 where R¹¹ is2-(N,N-dimethylamino)ethyl; R¹² is OAc; and R¹⁴ is H] is obtained bypurification of the crude product by use of HPLC.

[0567] Step 2

[0568] A solution of the compound [structure 66 where R¹¹ is2-(N,N-dimethylamino) ethyl; R¹² is OAc; and R¹⁴ is H] (1 mmol) preparedin the preceding reaction, the dihydropyridine [structure 75, where PGis H] (1 mmol), and diisopropylethylamine (0.2 mL) in DMF (5 mL) isstirred and warmed under an inert atmosphere. The progress of thereaction is followed by tic and when reaction is complete, the solutionis poured into aqueous 5% NaHCO₃ and the aqueous mixture is extractedwith methylene chloride. The organic extract solution is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired Formula I compound [structure 76, where R¹¹ is2-(N,N-dimethylamino)ethyl; R¹² is OAc; and R¹⁴ is H]compound isobtained by purification of the crude product by use of HPLC.

[0569] In similar manner, by replacing the benzothiazepine in the aboveexample with other ligands of structure 66 and/or by replacing thelinker molecule used to prepare 52 with other linker molecules, othercompounds of Formula I are prepared.

EXAMPLE 21

[0570] (see FIG. 18)

[0571] Preparation of a compound in which ligand, L₁, is linked directlyto ligand, L₂, where l₁ is the amlodipine moiety and L₂ is the diltiazemmoiety and whe re R¹H.

[0572] Method A

[0573] A mixture of NaH (1.1 mmol) and DMF (1 mL) is prepared under aninert atmosphere in a flask equipped with a s tirring b ar and a dryingtube. To this is added a solution of the benzothiazepine (structure 41where R¹² is OAc; R¹³ is methyl; and R¹⁴ is H) (1 mmol) in dry DMF (3mL) followed by 1,2-dibromoethane (10 mmol). The resulting mixture isstirred and the course of the reaction is followed by thin layerchromatography. After reaction occurs, the reaction is quenched withcold dilute aq. Na₂CO₃ and extracted with methylene chloride. Theorganic layer is dried (Na₂SO₄), filtered and concentrated under reducedpressure to give the crude product. The desired compound (structure 88)is obtained by purification of the crude product by use of HPLC.

[0574] Step 2

[0575] A solution of the compound (structure 88) (1 mmol) prepared inthe preceding reaction, N-BOC-amlodipine [structure 87, where PG ist-butyloxycarbonyl (BOC)] (1 mmol), and diisopropylethylamine (0.2 mL)in DMF (5 mL) is stirred and warmed under an inert atmosphere. Theprogress of the reaction is followed by tic and when reaction iscomplete, the solution is poured into aqueous 5% NaHCO₃ and the aqueousmixture is extracted with methylene chloride. The organic extractsolution is dried (Na₂SO₄), filtered and concentrated under reducedpressure to give the crude product. The desired compound (structure 89)is obtained by purification of the crude product by use of HPLC.

[0576] Step 3

[0577] A solution of the product (structure 89) from the precedingreaction and trifluoroacetic acid (3 mL) in CH₂Cl₂ (5 mL) is stirred atroom temperature. The progress of the reaction is followed by tic. Afterreaction occurs, more CH₂Cl₂ is added and the solution is washed withaqueous Na₂CO₃ and with H₂O. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound (structure 90) is obtained by purificationof the crude product with the use of HPLC.

[0578] Method B

[0579] A solution of the compound (structure 88) (1 mmol) prepared inthe preceding reaction, amlodipine (structure 87, where PG is H) (1mmol), and diisopropylethylamine (0.2 mL) in DMF (5 mL) is stirred andwarmed under an inert atmosphere. The progress of the reaction isfollowed by tic and when reaction is complete, the solution is pouredinto aqueous 5% NaHCO₃ and the aqueous mixture is extracted withmethylene chloride. The organic extract solution is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound (structure 90) is obtained by purificationof the crude product by use of HPLC.

EXAMPLE 22

[0580] (see FIG. 19)

[0581] Preparation of a compound in which ligand, L₁, is linked directlyto ligand, L₂, where L₁ is the amlodipine moiety and L₂ is the diltiazemmoiety

[0582] Method A

[0583] Step 1

[0584] A solution of compound (structure 89) (1 mmol) andparaformaldehyde (2 mmols) in methanol (4 mL) is stirred and isacidified with acetic acid to pH 6.6 (pH meter) under a nitrogenatmosphere. Sodium cyanoborohydride (1.1 mmol) is then added andstirring is continued. The course of the reaction is followed by thinlayer chromatography. After reaction occurs, the reaction solution isquenched in water and the pH of the aqueous mixture is adjusted togreater than 10 with aqueous NaOH. The mixture is extracted with ether,the organic extracts are washed with half-saturated saline, dried(Na₂SO₄), filtered and concentrated under reduced pressure to give thecrude product. The desired compound (structure 91) is obtained bypurification of the crude product by use of HPLC.

[0585] Step 2

[0586] A solution of the product (structure 91) from the precedingreaction and trifluoroacetic acid (3 mL) in CH₂Cl₂ (5 mL) is stirred atroom temperature. The progress of the reaction is followed by tic. Afterreaction occurs, more CH₂Cl₂ is added and the solution is washed withaqueous Na₂CO₃ and with H₂O. The organic layer is dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. The desired compound (structure 92) is obtained by purificationof the crude product with the use of HPLC.

[0587] Method B

[0588] Step 1

[0589] A solution of the compound (structure 89) (1 mmol) andparaformaldehyde (5 mmol) in ethanol (5 mL) is stirred with 10% Pd-C (20mg) under a hydrogen atmosphere. The progress of the reaction isfollowed by tic. After reaction occurs, the mixture is filtered throughCelite, the filter pad is washed with ethanol, and the filtrates areconcentrated under reduced pressure. The desired compound (structure 91)is obtained by purification of the crude product with the use of HPLCand is converted to structure 92 as described above in Method A.

[0590] (Method C)

[0591] A solution of the compound (structure 89) (1 mmol) in ether (5mL) is added slowly to a vigorously stirred mixture of a solution methyliodide (1 mmol) in ether (5 mL) and a solution of Na₂CO₃ in H₂0 (1 mL).The progress of the reaction is followed by tic. After reaction iscomplete, the mixture is washed with additional aqueous Na₂CO₃ and withH₂O, the organic layer is dried (Na₂SO₄), filtered and concentratedunder reduced pressure to give the crude product. The desired compound(structure 91) is obtained by purification of the crude product with theuse of HPLC. The compound (structure 91) is converted to structure 92 asdescribed above in Method A.

[0592] (Method D)

[0593] A solution of the compound (structure 90) (1 mmol) andparaformaldehyde (5 mmol) in ethanol (5 mL) is stirred with 10% Pd-C (20mg) under a hydrogen atmosphere. The progress of the reaction isfollowed by tic. After reaction occurs, the mixture is filtered throughCelite, the filter pad is washed with ethanol, and the filtrates areconcentrated under reduced pressure. The desired compound (structure 92)is obtained by purification of the crude product with the use of HPLC.

EXAMPLE 23

[0594] (see FIG. 20)

[0595] Preparation of a compound in which ligand, L₁, is linked directlyto ligand, L₂, where L₁ is the amlodipine moiety and L₂ is the diltiazemmoiety and where R=H.

[0596] Step 1

[0597] A mixture of ethanolamine (0.1 mol), di-tert-butylcarbonate (0.15mol), dioxane (50 mL) and aq. 2 N NaOH (25 mL) is stirred at RT for 24hr. The dioxane is removed by evaporation under reduced pressure. Water(50 mL) is added to the aqueous mixture and the mixture is extractedwith CH₂Cl₂ (4×25 mL). The combined organic layers are dried (Na₂SO₄),filtered and concentrated under reduced pressure to give the crudeproduct. Pure N-BOC-ethanolamine is obtained by purification of thecrude product with the use of flash chromatography over silica gel.

[0598] Step 2

[0599] t-Butyldimethylsilyl chloride(0.4 mol) is added to a solution ofN-BOC-ethanolamine (0.1 mole) and imidazole (0.1 mol) in dry pyridine(75 mL) and the resulting solution is stirred at RT. The progress of thereaction is followed by tic. When reaction is complete, water (5 mL) isadded to the solution which is then concentrated by evaporation underreduced pressure (>25 mm Hg, 30° C.). The residue is dissolved in EtOAcand the solution is extracted with saturated aq. CuSO₄ to removeresidual pyridine. The EtOAc solution is washed with water, dried(Na₂SO₄), filtered and concentrated under reduced pressure to give thecrude product. The pure product, N-BOC-ethanolamine-O-TBDMS, is obtainedby purification of the crude product by flash chromatography over silicagel.

[0600] Step 3

[0601] A solution of N-BOC-ethanolamine-O-TBDMS (0.05 mol) in dry DMF (3mL) is added dropwise to a stirred mixture of NaH (0.05 mol) and dry DMF(10 mL) under an inert atmosphere. The resulting mixture is stirred for1 hr and then is tranferred by cannulation to a stirred solution of1,2-dibromoethane (0.3 mol) in dry DMF (10 mL). The resulting solutionis stirred and the progress of the reaction is followed by tic. Afterreaction occurs, the reaction solution is quenched with aqueous 5%NaHCO₃ (100 mL) and brine (100 mL). The mixture is extracted with CH₂Cl₂(4×25 mL) and the combined organic extracts are back-washed with water(3×). The organic layer is dried (Na₂SO₄), filtered and concentratedunder reduced pressure to give the crude product. PureN-BOC-N-(2-bromoethyl) ethanolamine-O-TBDMS is obtained by purificationof the crude product with the use of flash chromatography over silicagel.

[0602] Step 4

[0603] A mixture of NaH (1.1 mmol) and dry DMF (1 mL) is prepared underan inert atmosphere in a flask equipped with a stirring bar and a dryingtube. To this is added a solution of the benzothiazepine (structure 41where R¹² is OAc; R¹³ is methyl; and R¹⁴ is H) (1 mmol) in dry DMF (3mL). Then a solution of N-BOC-N-(2-bromoethyl)ethanolamine-O-TBDMS (1mmol) in dry DMF (2 mL) is added and the resulting mixture is stirredand monitored for reaction by tic. After reaction occurs, the reactionsolution is quenched with aqueous 5% NaHCO₃ (25 mL) and brine (25 mL).The mixture is extracted with CH₂Cl₂ (4×20 mL) and the combined organicextracts are back-washed with water (3×). The organic layer is dried(Na₂SO₄), filtered and concentrated under reduced pressure to give thecrude product. The desired compound (structure 93) is obtained bypurification of the crude product with the use of HPLC.

[0604] Step 5

[0605] A solution of the product (structure 93) (1 mmol) from thepreceding reaction and Et₃N-(HF)₃ in MeCN (5 mL) is stirred at roomtemperature. After reaction occurs as detected by tic, the solution isdiluted with EtOAc and then washed with water-brine. The organic layeris dried (Na₂SO₄), filtered and concentrated under reduced pressure togive the crude product. The desired compound (structure 94) is obtainedby purification of the crude product with the use of HPLC.

[0606] Step 6

[0607] A mixture of NaH (2.1 mmol) and dry DMF (1 mL) is prepared underan inert atmosphere in a flask equipped with a stirring bar and a dryingtube. To this is added a solution of the compound (structure 94)prepared in the preceding reaction (1 mmol) in dry DMF (3 mL). Then asolution of the 1,4-dihydropyridine 26 (where PG=H) (1 mmol) in dry DMF(2 mL) is added and the resulting mixture is stirred and monitored forreaction by tic. After reaction occurs, the reaction solution isquenched water (25 mL) and brine (25 mL). The mixture is extracted withCH₂Cl₂ (4×20 mL) and the combined organic extracts are back-washed withwater (3×). The organic layer is dried (Na₂SO₄), filtered andconcentrated under reduced pressure to give the crude product. Thedesired compound (structure 89, where PG=H; R=BOC) is obtained bypurification of the crude product with the use of HPLC.

[0608] Step 7

[0609] A solution of the product (structure 89, PG=H; R=BOC) from thepreceding reaction and trifluoroacetic acid (3 mL) in CH₂Cl₂ (5 mL) isstirred at room temperature. The progress of the reaction is followed bytic. After reaction occurs, more CH₂Cl₂ is added and the solution iswashed with aqueous Na₂CO₃ and with H₂O. The organic layer is dried(Na₂SO₄), filtered and concentrated under reduced pressure to give thecrude product. The desired Formula I compound (structure 90) is obtainedby purification of the crude product with the use of HPLC.

[0610] While the present invention has been described with reference tothe specific embodiments thereof, it should be understood by thoseskilled in the art that various changes may be made and equivalents maybe substituted without departing from the true spirit and scope of theinvention. In addition, many modifications may be made to adapt aparticular situation, material, composition of matter, process, processstep or steps, to the objective spirit and scope of the presentinvention. All such modifications are intended to be within the scope ofthe claims appended hereto.

[0611] All of the publications, patent applications and patents cited inthis application are herein incorporated by reference in their entiretyto the same extent as if each individual publication, patent applicationor patent was specifically and individually indicated to be incorporatedby reference in its entirety. Multibinding Calcium: Channel AntagonistTable Cmpd Subunit 1 Linker Subunit 2 1 A

A 2 A

A 3 A

A 4 A

A 5 A

A 6 A

A 7 A

A 8 A

A 9 A

A 10 A

A 11 A

A 12 A

A 13 A

A 14 A

A 15 A

A 16 A

A 17 A

A 18 A

A 19 A

A 20 A

A 21 A

A 22 A

A 23 A

A 24 A

A 25 A

A 26 A

A 27 A

A 28 A

A 29 A

A 30 A

A 31 A

A 32 A

A 33 A

A 33 A

A 34 A

A 35 A

A 36 A

A 37 A

B 38 A

B 39 A

B 41 A

B 42 A

B 43 A

B 44 A

B 45 A

B 46 A

B 47 A

B 48 A

B 49 A

B 50 A

B 51 A

B 52 A

B 53 A

B 54 A

B 55 A

B 56 A

B 57 A

B 58 A

B 59 A

B 60 A

B 61 A

B 62 A

B 63 A

B 64 A

B 65 A

B 66 A

B 67 A

B 68 A

B 69 A

B 70 A

B 71 A

B 72 A

B 73 A

B 75 A

C 76 A

C 77 A

C 78 A

C 79 A

C 80 A

C 81 A

C 82 A

C 83 A

C 84 A

C 85 A

C 86 A

C 87 A

C 88 A

C 89 A

C 90 A

C 91 A

C 92 A

C 93 A

C 94 A

C 95 A

C 96 A

C 97 A

C 98 A

C 99 A

C 100 A

C 101 A

C 102 A

C 103 A

C 104 A

C 105 A

C 106 A

C 107 A

C 108 A

C 109 A

C 110 A

C 111 A

C 112 A

D 113 A

D 114 A

D 115 A

D 116 A

D 117 A

D 118 A

D 119 A

D 120 A

D 121 A

D 122 A

D 123 A

D 124 A

D 125 A

D 126 A

D 127 A

D 128 A

D 129 A

D 130 A

D 131 A

D 132 A

D 133 A

D 134 A

D 135 A

D 136 A

D 137 A

D 138 A

D 139 A

D 140 A

D 141 A

D 142 A

D 143 A

D 144 A

D 145 A

D 146 A

D 147 A

D 148 A

D 149 A

E 150 A

E 151 A

E 152 A

E 153 A

E 154 A

E 155 A

E 156 A

E 157 A

E 158 A

E 159 A

E 160 A

E 161 A

E 162 A

E 163 A

E 164 A

E 165 A

E 166 A

E 167 A

E 168 A

E 169 A

E 170 A

E 171 A

E 172 A

E 173 A

E 174 A

E 175 A

E 176 A

E 177 A

E 178 A

E 179 A

E 180 A

E 181 A

E 182 A

E 183 A

E 184 A

E 185 A

E 186 A

H 187 A

H 188 A

H 189 A

H 190 A

H 191 A

H 192 A

H 193 A

H 194 A

H 195 A

H 196 A

H 197 A

H 198 A

H 199 A

H 200 A

H 201 A

H 202 A

H 203 A

H 204 A

H 205 A

H 206 A

H 207 A

H 208 A

H 209 A

H 210 A

H 211 A

H 212 A

H 213 A

H 214 A

H 215 A

H 216 A

H 217 A

H 218 A

H 219 A

H 220 A

H 221 A

H 222 A

H 223 B

C 224 B

C 225 B

C 226 B

C 227 B

C 228 B

C 229 B

C 230 B

C 231 B

C 232 B

C 233 B

C 234 B

C 235 B

C 236 B

C 237 B

C 238 B

C 239 B

C 249 B

C 250 B

C 251 B

C 252 B

C 253 B

C 254 B

C 255 B

C 256 B

C 257 B

C 258 B

C 259 B

C 260 B

C 261 B

C 262 B

C 263 B

C 264 B

C 265 B

C 266 B

C 267 B

C 268 B

C 269 B

D 270 B

D 271 B

D 272 B

D 273 B

D 274 B

D 275 B

D 276 B

D 277 B

D 278 B

D 279 B

D 280 B

D 281 B

D 282 B

D 283 B

D 284 B

D 285 B

D 286 B

D 287 B

D 288 B

D 289 B

D 290 B

D 291 B

D 292 B

D 293 B

D 294 B

D 295 B

D 296 B

D 297 B

D 298 B

D 299 B

D 300 B

D 301 B

D 302 B

D 303 B

D 304 B

D 305 B

D 306 B

E 307 B

E 308 B

E 309 B

E 310 B

E 311 B

E 312 B

E 313 B

E 314 B

E 315 B

E 316 B

E 317 B

E 318 B

E 319 B

E 320 B

E 321 B

E 322 B

E 323 B

E 324 B

E 325 B

E 326 B

E 327 B

E 328 B

E 329 B

E 330 B

E 331 B

E 332 B

E 333 B

E 334 B

E 335 B

E 336 B

E 337 B

E 338 B

E 339 B

E 340 B

E 341 B

E 342 B

E 343 B

H 344 B

H 345 B

H 346 B

H 347 B

H 348 B

H 349 B

H 350 B

H 351 B

H 352 B

H 353 B

H 354 B

H 355 B

H 356 B

H 357 B

H 358 B

H 359 B

H 360 B

H 361 B

H 362 B

H 363 B

H 364 B

H 365 B

H 366 B

H 367 B

H 368 B

H 369 B

H 370 B

H 371 B

H 372 B

H 373 B

H 374 B

H 375 B

H 376 B

H 377 B

H 378 B

H 379 B

H 380 C

H 381 C

H 382 C

H 383 C

H 384 C

H 385 C

H 386 C

H 387 C

H 388 C

H 389 C

H 390 C

H 391 C

H 392 C

H 393 C

H 394 C

H 395 C

H 396 C

H 397 C

H 398 C

H 399 C

H 400 C

H 401 C

H 402 C

H 403 C

H 404 C

H 405 C

H 406 C

H 407 C

H 408 C

H 409 C

H 410 C

H 411 C

H 412 C

H 413 C

H 414 C

H 415 C

H 416 C

H 417 D

H 418 D

H 419 D

H 420 D

H 421 D

H 422 D

H 423 D

H 424 D

H 425 D

H 426 D

H 427 D

H 428 D

H 429 D

H 430 D

H 431 D

H 432 D

H 433 D

H 434 D

H 435 D

H 436 D

H 437 D

H 438 D

H 439 D

H 440 D

H 441 D

H 442 D

H 443 D

H 444 D

H 445 D

H 446 D

H 447 D

H 448 D

H 449 D

H 450 D

H 451 D

H 452 D

H 453 D

H 454 E

H 455 E

H 456 E

H 457 E

H 458 E

H 459 E

H 460 E

H 461 E

H 462 E

H 463 E

H 464 E

H 465 E

H 466 E

H 467 E

H 468 E

H 469 E

H 470 E

H 471 E

H 472 E

H 473 E

H 474 E

H 475 E

H 476 E

H 477 E

H 478 E

H 479 E

H 480 E

H 481 E

H 482 E

H 483 E

H 484 E

H 485 E

H 486 E

H 487 E

H 488 E

H 489 E

H 490 E

H 491 F None A 492 F

A 493 F

A 494 F

A 495 F

A 496 F

A 497 F

A 498 F

A 499 F

A 500 F

A 501 F

A 502 F

A 503 F

A 504 F

A 505 F

A 506 F

A 507 F

A 508 F

A 509 F

A 510 F

A 511 F

A 512 F

A 513 F

A 514 F

A 515 F

A 516 F

A 517 F

A 518 F

A 491 F None C 492 F

C 493 F

C 494 F

C 495 F

C 496 F

C 497 F

C 498 F

C 499 F

C 500 F

C 501 F

C 502 F

C 503 F

C 504 F

C 505 F

C 506 F

C 507 F

C 508 F

C 509 F

C 510 F

C 511 F

C 512 F

C 513 F

C 514 F

C 515 F

C 516 F

C 517 F

C 518 F

C 519 F None D 520 F

D 521 F

D 522 F

D 523 F

D 524 F

D 525 F

D 526 F

D 527 F

D 528 F

D 529 F

D 530 F

D 531 F

D 532 F

D 533 F

D 534 F

D 535 F

D 536 F

D 537 F

D 538 F

D 539 F

D 540 F

D 541 F

D 542 F

D 543 F

D 544 F

D 545 F

D 546 F

D 547 F

E 548 F

E 549 F

E 550 F

E 551 F

E 552 F

E 553 F

E 554 F

E 555 F

E 556 F

E 557 F

E 558 F

E 559 F

E 560 F

E 561 F

E 562 F

E 563 F

E 564 F

E 565 F

E 566 F

E 567 F

E 568 F

E 569 F

E 570 F

E 571 F

E 572 F

E 573 F

E 574 F None H 575 F

H 576 F

H 577 F

H 578 F

H 579 F

H 580 F

H 581 F

H 582 F

H 583 F

H 584 F

H 585 F

H 586 F

H 587 F

H 588 F

H 589 F

H 590 F

H 591 F

H 592 F

H 593 F

H 594 F

H 595 F

H 596 F

H 597 F

H 598 F

H 599 F

H 600 F

H 601 F

H 602 G None A 603 G

A 604 G

A 605 G

A 606 G

A 607 G

A 608 G

A 609 G

A 610 G

A 611 G

A 612 G

A 613 G

A 614 G

A 615 G

A 616 G

A 617 G

A 618 G

A 619 G

A 620 G

A 621 G

A 622 G

A 623 G

A 624 G

A 625 G

A 626 G

A 627 G

A 628 G

A 629 G

A 630 G None C 631 G

C 632 G

C 633 G

C 634 G

C 635 G

C 636 G

C 637 G

C 638 G

C 639 G

C 640 G

C 641 G

C 642 G

C 643 G

C 644 G

C 645 G

C 646 G

C 647 G

C 648 G

C 649 G

C 650 G

C 651 G

C 652 G

C 653 G

C 654 G

C 655 G

C 656 G

C 657 G

C 658 G None D 659 G

D 660 G

D 661 G

D 662 G

D 663 G

D 664 G

D 665 G

D 666 G

D 667 G

D 668 G

D 669 G

D 670 G

D 671 G

D 672 G

D 673 G

D 674 G

D 675 G

D 676 G

D 677 G

D 678 G

D 679 G

D 680 G

D 681 G

D 682 G

D 683 G

D 684 G

D 685 G

D 686 G None E 687 G

E 688 G

E 689 G

E 690 G

E 691 G

E 692 G

E 693 G

E 694 G

E 695 G

E 696 G

E 697 G

E 698 G

E 699 G

E 700 G

E 701 G

E 702 G

E 703 G

E 704 G

E 705 G

E 706 G

E 707 G

E 708 G

E 709 G

E 710 G

E 711 G

E 712 G

E 713 G

E 714 G None H 715 G

H 716 G

H 717 G

H 718 G

H 719 G

H 720 G

H 721 G

H 722 G

H 723 G

H 724 G

H 725 G

H 726 G

H 727 G

H 728 G

H 729 G

H 730 G

H 731 G

H 732 G

H 733 G

H 734 G

H 735 G

H 736 G

H 737 G

H 738 G

H 739 G

H 740 G

H 741 G

H

What is claimed is:
 1. A multibinding compound comprising 2 to 10ligands which may be the same or different and which are covalentlyattached to a linker or linkers, which may be the same or different,each of said ligands comprising a ligand domain capable of binding to aCa⁺⁺ channel.
 2. A multibinding compound according to claim 1, whereinthe ligands are selected from A-53930A, AE-0047, AGN-190604, AGN-190744,AH-1058, AHR-12742, AHR-16303B, AHR-16462B, AIT-10, AIT-111, AJ-3941,AM-336, amlopidine (including S-(−), R-(+), and racemic), anipamil,AP-1067, aranidipine, atosiban, azelnidipine, barnidipine, Bay-t-7207,Bay-y-5959, Bay-z-4406, BBR-2160, belfosdil, B111-890-CL, bisaramil,BMS-181102, BMS-188107, BMY-43011, BRL-32872, buflomedil, CD-349,CD-832, CERM-12816, CGP-28932, cilnidipine, clentiazem, clevidipine,CNS-1067, CNS-1237, CNS-2103, CP-060S, CPC-301, CPC-317, CPU-86017,D-2024, darodipine, DHP-218, diltiazem, diperdipine, dopropidil,dotarazine, dronedarone, DTZ-323, E-047/1, efonidipine, EGIS-7229,elgodipine, emopamil, etomoxir, F-0401, fantofarone, fasudil, FCE-24265,FCE-26262, FCE-27335, FCE-27892, FCE-28718, felodipine, FPL-64176,FR-172516, FRG-8701, furnidipine, GS-386, iganidipine, ipenoxazone,isradipine, JTV-591, KP-840, KT-362, L-366682, lacidipine, LAS-0538,LCB-2514, lemildipine, lercanidipine, leualacin, lifarizine, LOE-908,lomerizine, lubeluzole, LY-042826, manidipine, McN-6186, mibefradil,monatepil , MR-14134, N-3601, NCC-1048, nefiracetam, nexopamil,nifedipine, nifedipine, Nifelan, nilvadipine, nimodipine, NNC-09-0026,NPS-568, NS-638, NS-649, NS-696, NS-7, OPC-8490, Org-13061, Org-30029,oxodipine, P-5, palonidipine, PCA-50922, PCA-50938, PCA-50941,PD-029361, PD-157667, PD-158143, PD-176078, pranidipine, QX-314,ranolazine, RHG-2716, RingCap, Ro-1 1-2933, RS-5773, RU-43945,RWJ-22108, RWJ-22726, RWJ-29009, RWJ-37868, S-12968, S-2150, S-312-d,SANK-71996, SB-201823, SB-206284A, SB-23736, SD-3212, semotiadil,SIB-1281, siratiazem, SKF-45675, SKF-96365, SKT-M-26, SL-34.0829,SL-87.0495, SM-6586, SNX-124, SNX-236, SNX-239, SNX-325, SNX-482,SQ-31727, SQ-33351, SQ-34399, SR-33805, TA-993, tamolarizine, TDN-345,temiverine, terodiline, TH9229, TN-871, U-88999, U-92032, U-92798,UCL-1439, UK-1 656, UK-55444, UK-56593, UK-84149, verapamil, Verelan,vexibinol, VUF-8929, WAY-141520, XB-513, XT-044, Y-22516, YH-334,YM-1615-4, YM-430, Z-6568, zatebradine, ziconotide, and ZM-224832.
 3. Amultibinding compound according to claim 2, wherein the ligands areselected from the group consisting of verapamil, diltiazem, benziazemclentiazem, nicardipine, nifedipine, nilvadipine, nitredipine,nimodipine, isradipine, lacidipine, amlodipine, nisoldipine, isradipine,mibefrodil, amlodipine, felodipine, nimodipine, bepridil, SQ 32,910 andSQ 32,428.
 4. A multibinding compound represented by Formula I:(L)_(p)(X)_(q)  I where each L is a ligand that may be the same ordifferent at each occurrence; X is a linker that may be the same ordifferent at each occurrence; p is an integer of from 2 to 10; and q isan integer of from 1 to 20; wherein each of said ligands comprises aligand domain capable of binding to a Ca⁺⁺ channel.
 5. A multibindingcompound according to claim 4, wherein q is less than p.
 6. Amultibinding compound according to claim 4, wherein the ligands areselected from A-53930A, AE-0047, AGN-190604, AGN-190744, AH-1058,AHR-12742, AHR-16303B, AHR-16462B, AIT-1 10, AIT-111, AJ-3941, AM-336,amlopidine (including S-(−), R-(+), and racemic), anipamil, AP-1067,aranidipine, atosiban, azelnidipine, barnidipine, Bay-t-7207,Bay-y-5959, Bay-z-4406, BBR-2160, belfosdil, BIII-890-CL, bisaramil,BMS-181102, BMS-188107, BMY-43011, BRL-32872, buflomedil, CD-349,CD-832, CERM-12816, CGP-28932, cilnidipine, clentiazem, clevidipine,CNS-1067, CNS-1237, CNS-2103, CP-060S, CPC-301, CPC-317, CPU-86017,D-2024, darodipine, DHP-218, diltiazem, diperdipine, dopropidil,dotarazine, dronedarone, DTZ-323, E-047/1, efonidipine, EGIS-7229,elgodipine, emopamil, etomoxir, F-0401, fantofarone, fasudil, FCE-24265,FCE-26262, FCE-27335, FCE-27892, FCE-28718, felodipine, FPL-64176,FR-172516, FRG-8701, furnidipine, GS-386, iganidipine, ipenoxazone,isradipine, JTV-591, KP-840, KT-362, L-366682, lacidipine, LAS-0538,LCB-2514, lemildipine, lercanidipine, leualacin, lifarizine, LOE-908,lomerizine, lubeluzole, LY-042826, manidipine, McN-6186, mibefradil,monatepil , MR-14134, N-3601, NCC-10⁴⁸, nefiracetam, nexopamil,nifedipine, nifedipine, Nifelan, nilvadipine, nimodipine, NNC-09-0026,NPS-568, NS-638, NS-649, NS-696, NS-7, OPC-8490, Org-13061, Org-30029,oxodipine, P-5, palonidipine, PCA-50922, PCA-50938, PCA-50941,PD-029361, PD-157667, PD-1 58143, PD-1 76078, pranidipine, QX-314,ranolazine, RHG-2716, RingCap, Ro-11-2933, RS-5773, RU-43945, RWJ-22108,RWJ-22726, RWJ-29009, RWJ-37868, S-12968, S-2150, S-312-d, SANK-71996,SB-201823, SB-206284A, SB-23736, SD-3212, semotiadil, SIB-1281,siratiazem, SKF-45675, SKF-96365, SKT-M-26, SL-34.0829, SL-87.0495,SM-6586, SNX-124, SNX-236, SNX-239, SNX-325, SNX-482, SQ-31727,SQ-33351, SQ-34399, SR-33805, TA-993, tamolarizine, TDN-345, temiverine,terodiline, TH9229, TN-871, U-88999, U-92032, U-92798, UCL-1439, UK-1656, UK-55444, UK-56593, UK-84149, verapamil, Verelan, vexibinol,VUF-8929, WAY-141520, XB-513, XT-044, Y-22516, YH-334, YM-1615-4,YM-430, Z-6568, zatebradine, ziconotide, and ZM-224832.
 7. Amultibinding compound according to claim 6, wherein the ligands areselected from the group consisting of verapamil, diltiazem, benziazemclentiazem, nicardipine, nifedipine, nilvadipine, nitredipine,nimodipine, isradipine, lacidipine, amlodipine, nisoldipine, isradipine,mibefrodil, amlodipine, felodipine, nimodipine, bepridil, SQ 32,910 andSQ 32,428.
 8. The multibinding compound of claim 4, wherein the linkeris is represented by the following formula: —X′—Z—(Y′—Z)_(m)—Y″—Z—X′— inwhich: m is an integer of from 0 to 20; X′ at each separate occurrenceis —O—, —S—, —S(O)—, —S(O)₂—, —NR—, —N⁺ R R′—, —C(O)—, —C(O)O—,—C(O)NH—, —C(S), —C(S)O—, —C(S)NH— or a covalent bond, where R and R ateach separate occurrence are as defined below for R′and R″; Z is at eachseparate occurrence selected from alkylene, substituted alkylene,alkylalkoxy, cycloalkylene, substituted cycloalkylene, alkenylene,substituted alkenylene, alkynylene, substituted alkynylene,cycloalkenylene, substituted alkenylene, arylene, substituted arylene,heteroarylene, heterocyclene, substituted heterocyclene, crowncompounds, or a covalent bond; Y′ and Y′ at each separate occurrence areselected from —S—S— or a covalent bond;

in which: n is 0, 1 or 2; and R′ and R″ at each separate occurrence areselected from hydrogen, alkyl, substituted al kyl, cycloal kyl,substituted cycloal kyl, aIIkenyl, substituted alkenyl, alkynyl,substituted alkynyl, aryl, heteroaryl or heterocyclic.
 9. Themultibinding compound of claim 8 wherein p is 2 and q is
 1. 10. Apharmaceutical composition comprising a pharmaceutically acceptableexcipient and a therapeutically effective amount of one or moremultibindimg compounds, or pharmaceutically acceptable salts thereof,comprising 2 to 10 ligands which may be the same or different and whichare covalently attached to a linker or linkers, which may be the same ordifferent, each of said ligands comprising a ligand domain capable ofbinding to a Ca⁺⁺ channel of a cell mediating mammalian diseases orconditions, thereby modulating the diseases or conditions.
 11. Apharmaceutical composition according to claim 10, wherein the ligandsare selected from A-53930A, AE-0047, AGN-190604, AGN-190744, AH-1058,AHR-12742, AHR-16303B, AHR-16462B, AIT-110, AIT-111, AJ-3941, AM-336,amlopidine (including S-(−), R-(+), and racemic), anipamil, AP-1067,aranidipine, atosiban, azelnidipine, barnidipine, Bay-t-7207,Bay-y-5959, Bay-z-4406, BBR-2160, belfosdil, BIII-890-CL, bisaramil,BMS-181102, BMS-188107, BMY-43011, BRL-32872, buflomedil, CD-349,CD-832, CERM-12816, CGP-28932, cilnidipine, clentiazem, clevidipine,CNS-1067, CNS-1237, CNS-2103, CP-060S, CPC-301, CPC-317, CPU-86017,D-2024, darodipine, DHP-218, diltiazem, diperdipine, dopropidil,dotarazine, dronedarone, DTZ-323, E-047/1, efonidipine, EGIS-7229,elgodipine, emopamil, etomoxir, F-0401, fantofarone, fasudil, FCE-24265,FCE-26262, FCE-27335, FCE-27892, FCE-28718, felodipine, FPL-64176,FR-172516, FRG-8701, furnidipine, GS-386, iganidipine, ipenoxazone,isradipine, JTV-591, KP-840, KT-362, L-366682, lacidipine, LAS-0538,LCB-2514, lemildipine, lercanidipine, leualacin, lifarizine, LOE-908,lomerizine, lubeluzole, LY-042826, manidipine, McN-6186, mibefradil,monatepil, MR-14134, N-3601, NCC-1048, nefiracetam, nexopamil,nifedipine, nifedipine, Nifelan, nilvadipine, nimodipine, NNC-09-0026,NPS-568, NS-638, NS-649, NS-696, NS-7, OPC-8490, Org-13061, Org-30029,oxodipine, P-5, palonidipine, PCA-50922, PCA-50938, PCA-50941,PD-029361, PD-157667, PD-158143, PD-176078, pranidipine, QX-314,ranolazine, RHG-2716, RingCap, Ro-11-2933, RS-5773, RU-43945, RWJ-22108,RWJ-22726, RWJ-29009, RWJ-37868, S-12968, S-2150, S-312-d, SANK-71996,SB-201823, SB-206284A, SB-23736, SD-3212, semotiadil, SIB-1281,siratiazem, SKF-45675, SKF-96365, SKT-M-26, SL-34.0829, SL-87.0495,SM-6586, SNX-124, SNX-236, SNX-239, SNX-325, SNX-482, SQ-31727,SQ-33351, SQ-34399, SR-33805, TA-993, tamolarizine, TDN-345, temiverine,terodiline, TH9229, TN-871, U-88999, U-92032, U-92798, UCL-1439,UK-1656, UK-55444, UK-56593, UK-84149, verapamil, Verelan, vexibinol,VUF-8929, WAY-141520, XB-513, XT-044, Y-22516, YH-334, YM-1615-4,YM-430, Z-6568, zatebradine, ziconotide, and ZM-224832.
 12. Apharmaceutical composition according to claim 11, wherein the ligandsare selected from the group consisting of verapamil, diltiazem,benziazem clentiazem, nicardipine, nifedipine, nilvadipine, nitredipine,nimodipine, isradipine, lacidipine, amlodipine, nisoldipine, isradipine,mibefrodil, amlodipine, felodipine, nimodipine, bepridil, SQ 32,910 andSQ 32,428.
 13. A pharmaceutical composition comprising apharmaceutically acceptable excipient and a therapeutically effectiveamount of one or more multibinding compounds represented by Formula I,(L)_(p)(X)_(q)  I and pharmaceutically acceptable salts thereof, whereeach L is a ligand that may be the same or different at each occurrence;X is a linker that may be the same or different at each occurrence; p isan integer of from 2 to 10; and q is an integer of from 1 to 20; whereineach of said ligands comprises a ligand domain capable of binding to aCa⁺⁺ channel of a cell mediating mammalian diseases or conditions,thereby modulating the diseases or conditions.
 14. A pharmaceuticalcomposition according to claim 4, wherein the ligands are selected fromA-53930A, AE-0047, AGN-190604, AGN-190744, AH-1058, AHR-12742,AHR-16303B, AHR-16462B, AIT-i 10, AIT-111, AJ-3941, AM-336, amlopidine(including S-(−), R-(+), and racemic), anipamil, AP-1067, aranidipine,atosiban, azelnidipine, barnidipine, Bay-t-7207, Bay-y-5959, Bay-z-4406,BBR-2160, belfosdil, BIII-890-CL, bisaramil, BMS-181102, BMS-188107,BMY-43011, BRL-32872, buflomedil, CD-349, CD-832, CERM-12816, CGP-28932,cilnidipine, clentiazem, clevidipine, CNS-1067, CNS-1237, CNS-2103,CP-060S, CPC-301, CPC-317, CPU-86017, D-2024, darodipine, DHP-218,diltiazem, diperdipine, dopropidil, dotarazine, dronedarone, DTZ-323,E-047/1, efonidipine, EGIS-7229, elgodipine, emopamil, etomoxir, F-0401,fantofarone, fasudil, FCE-24265, FCE-26262, FCE-27335, FCE-27892,FCE-28718, felodipine, FPL-64176, FR-172516, FRG-8701, furnidipine,GS-386, iganidipine, ipenoxazone, isradipine, JTV-591, KP-840, KT-362,L-366682, lacidipine, LAS-0538, LCB-2514, lemildipine, lercanidipine,leualacin, lifarizine, LOE-908, lomerizine, lubeluzole, LY-042826,manidipine, McN-6186, mibefradil, monatepil, MR-14134, N-3601, NCC-1048,nefiracetam, nexopamil, nifedipine, nifedipine, Nifelan, nilvadipine,nimodipine, NNC-09-0026, NPS-568, NS-638, NS-649, NS-696, NS-7,OPC-8490, Org-13061, Org-30029, oxodipine, P-5, palonidipine, PCA-50922,PCA-50938, PCA-50941, PD-029361, PD-157667, PD-158143, PD-176078,pranidipine, QX-314, ranolazine, RHG-2716, RingCap, Ro-11-2933, RS-5773,RU-43945, RWJ-22108, RWJ-22726, RWJ-29009, RWJ-37868, S-12968, S-2150,S-312-d, SANK-71996, SB-201823, SB-206284A, SB-23736, SD-3212,semotiadil, SIB-1281, siratiazem, SKF-45675, SKF-96365, SKT-M-26,SL-34.0829, SL-87.0495, SM-6586, SNX-124, SNX-236, SNX-239, SNX-325,SNX-482, SQ-31727, SQ-33351, SQ-34399, SR-33805, TA-993, tamolarizine,TDN-345, temiverine, terodiline, TH9229, TN-871, U-88999, U-92032,U-92798, UCL-1439, UK-1656, UK-55444, UK-56593, UK-84149, verapamil,Verelan, vexibinol, VUF-8929, WAY-141520, XB-513, XT-044, Y-22516,YH-334, YM-1615-4, YM-430, Z-6568, zatebradine, ziconotide, andZM-224832.
 15. A pharmaceutical composition according to claim 6,wherein the ligands are selected from the group consisting of verapamil,diltiazem, benziazem clentiazem, nicardipine, nifedipine, nilvadipine,nitredipine, nimodipine, isradipine, lacidipine, amlodipine,nisoldipine, isradipine, mibefrodil, amlodipine, felodipine, nimodipine,bepridil, SQ 32,910 and SQ 32,428.
 16. A method for modulating theactivity of a Ca⁺⁺ channel in a biologic tissue, which method comprisescontacting a tissue having a Ca⁺⁺ channel with a multibinding compound,or a pharmaceutically acceptable salt thereof, under conditionssufficient to produce a change in the activity of the channel in saidtissue, wherein the multibinding compound comprises 2 to 10 ligandswhich may be the same or different and which are covalently attached toa linker or linkers, which may be the same or different, each of saidligands comprising a ligand domain capable of binding to a Ca⁺⁺ channel.17. A method according to claim 16, wherein the ligands are selectedfrom A-53930A, AE-0047, AGN-190604, AGN-190744, AH-1058, AHR-12742,AHR-16303B, AHR-16462B, AIT-110, AIT-111, AJ-3941, AM-336, amlopidine(including S-(−), R-(+), and racemic), anipamil, AP-1067, aranidipine,atosiban, azelnidipine, barnidipine, Bay-t-7207, Bay-y-5959, Bay-z-4406,BBR-2160, belfosdil, B111-890-CL, bisaramil, BMS-181102, BMS-1 88107,BMY-43011, BRL-32872, buflomedil, CD-349, CD-832, CERM-12816, CGP-28932,cilnidipine, clentiazem, clevidipine, CNS-1067, CNS-1237, CNS-2103,CP-060S, CPC-301, CPC-317, CPU-86017, D-2024, darodipine, DHP-218,diltiazem, diperdipine, dopropidil, dotarazine, dronedarone, DTZ-323,E-047/1, efonidipine, EGIS-7229, elgodipine, emopamil, etomoxir, F-0401,fantofarone, fasudil, FCE-24265, FCE-26262, FCE-27335, FCE-27892,FCE-28718, felodipine, FPL-64176, FR-172516, FRG-8701, furnidipine,GS-386, iganidipine, ipenoxazone, isradipine, JTV-591, KP-840, KT-362,L-366682, lacidipine, LAS-0538, LCB-2514, lemildipine, lercanidipine,leualacin, lifarizine, LOE-908, lomerizine, lubeluzole, LY-042826,manidipine, McN-6186, mibefradil, monatepil, MR-14134, N-3601, NCC-1048,nefiracetam, nexopamil, nifedipine, nifedipine, Nifelan, nilvadipine,nimodipine, NNC-09-0026, NPS-568, NS-638, NS-649, NS-696, NS-7,OPC-8490, Org-13061, Org-30029, oxodipine, P-5, palonidipine, PCA-50922,PCA-50938, PCA-50941, PD-029361, PD-157667, PD-1 58143, PD-1 76078,pranidipine, QX-314, ranolazine, RHG-2716, RingCap, Ro-11-2933, RS-5773,RU-43945, RWJ-22108, RWJ-22726, RWJ-29009, RWJ-37868, S-12968, S-2150,S-312-d, SANK-71996, SB-201823, SB-206284A, SB-23736, SD-3212,semotiadil, SIB-1281, siratiazem, SKF-45675, SKF-96365, SKT-M-26,SL-34.0829, SL-87.0495, SM-6586, SNX-124, SNX-236, SNX-239, SNX-325,SNX-482, SQ-31727, SQ-33351, SQ-34399, SR-33805, TA-993, tamolarizine,TDN-345, temiverine, terodiline, TH9229, TN-871, U-88999, U-92032,U-92798, UCL-1439, UK-1656, UK-55444, UK-56593, UK-84149, verapamil,Verelan, vexibinol, VUF-8929, WAY-141520, XB-513, XT-044, Y-22516,YH-334, YM-1615-4, YM-430, Z-6568, zatebradine, ziconotide, andZM-224832.
 18. A method according to claim 17, wherein the ligands areselected from the group consisting of verapamil, diltiazem, benziazemclentiazem, nicardipine, nifedipine, nilvadipine, nitredipine,nimodipine, isradipine, lacidipine, amlodipine, nisoldipine, isradipine,mibefrodil, amlodipine, felodipine, nimodipine, bepridil, SQ 32,910 andSQ 32,428.
 19. A method for treating a disease or condition in a mammalresulting from an activity of a Ca⁺⁺ channel, which method comprisesadministering to said mammal a therapeutically effective amount of apharmaceutical composition comprising a pharmaceutically acceptableexcipient and one or more multibinding compounds, or pharmaceuticallyacceptable salts thereof, comprising 2 to 10 ligands which may be thesame or different and which are covalently attached to a linker orlinkers, which may be the same or different, each of said ligandscomprising a ligand domain capable of binding to a Ca⁺⁺ channel of acell mediating mammalian diseases or conditions.
 20. A method accordingto claim 19, wherein the ligands are selected from A-53930A, AE-0047,AGN-190604, AGN-190744, AH-1058, AHR-12742, AHR-1 6303B, AHR-1 6462B,AIT-110, AIT-111, AJ-3941, AM-336, amlopidine (including S-(−), R-(+),and racemic), anipamil, AP-1067, aranidipine, atosiban, azelnidipine,barnidipine, Bay-t-7207, Bay-y-5959, Bay-z-4406, BBR-2160, belfosdil,BIII-890-CL, bisaramil, BMS-181102, BMS-1 88107, BMY-43011, BRL-32872,buflomedil, CD-349, CD-832, CERM-12816, CGP-28932, cilnidipine,clentiazem, clevidipine, CNS-1067, CNS-1237, CNS-2103, CP-060S, CPC-301,CPC-317, CPU-86017, D-2024, darodipine, DHP-218, diltiazem, diperdipine,dopropidil, dotarazine, dronedarone, DTZ-323, E-047/1, efonidipine,EGIS-7229, elgodipine, emopamil, etomoxir, F-0401, fantofarone, fasudil,FCE-24265, FCE-26262, FCE-27335, FCE-27892, FCE-28718, felodipine,FPL-64176, FR-172516, FRG-8701, furnidipine, GS-386, iganidipine,ipenoxazone, isradipine, JTV-591, KP-840, KT-362, L-366682, lacidipine,LAS-0538, LCB-2514, lemildipine, lercanidipine, leualacin, lifarizine,LOE-908, lomerizine, lubeluzole, LY-042826, manidipine, McN-6186,mibefradil, monatepil , MR-14134, N-3601, NCC-1048, nefiracetam,nexopamil, nifedipine, nifedipine, Nifelan, nilvadipine, nimodipine,NNC-09-0026, NPS-568, NS-638, NS-649, NS-696, NS-7, OPC-8490, Org-13061, Org-30029, oxodipine, P-5, palonidipine, PCA-50922, PCA-50938,PCA-50941, PD-029361, PD-157667, PD-158143, PD-176078, pranidipine,QX-314, ranolazine, RHG-2716, RingCap, Ro-11-2933, RS-5773, RU-43945,RWJ-22108, RWJ-22726, RWJ-29009, RWJ-37868, S-12968, S-2150, S-312-d,SANK-71996, SB-201823, SB-206284A, SB-23736, SD-3212, semotiadil,SIB-1281, siratiazem, SKF-45675, SKF-96365, SKT-M-26, SL-34.0829,SL-87.0495, SM-6586, SNX-124, SNX-236, SNX-239, SNX-325, SNX-482,SQ-31727, SQ-33351, SQ-34399, SR-33805, TA-993, tamolarizine, TDN-345,temiverine, terodiline, TH9229, TN-871, U-88999, U-92032, U-92798,UCL-1439, UK-1656, UK-55444, UK-56593, UK-84149, verapamil, Verelan,vexibinol, VUF-8929, WAY-141520, XB-513, XT-044, Y-22516, YH-334,YM-1615-4, YM-430, Z-6568, zatebradine, ziconotide, and ZM-224832.
 21. Amethod according to claim 20, wherein the ligands are selected from thegroup consisting of verapamil, diltiazem, benziazem clentiazem,nicardipine, nifedipine, nilvadipine, nitredipine, nimodipine,isradipine, lacidipine, amlodipine, nisoldipine, isradipine, mibefrodil,amlodipine, felodipine, nimodipine, bepridil, SQ 32,910 and SQ 32,428.22. A method for treating a disease or condition in a mammal resultingfrom an activity of a Ca⁺⁺ channel, which method comprises administeringto said mammal a therapeutically effective amount of a pharmaceuticalcomposition comprising a pharmaceutically acceptable excipient and oneor more multibinding compounds represented by Formula I,(L)_(p)(X)_(q)  I and pharmaceutically acceptable salts thereof, whereeach L is a ligand that may be the same or different at each occurrence;X is a linker that may be the same or different at each occurrence; p isan integer of from 2 to 10; and q is an integer of from 1 to 20; whereineach of said ligands comprises a ligand domain capable of binding to aCa⁺⁺ channel of a cell mediating mammalian diseases or conditions.
 23. Amethod according to claim 22, wherein the ligands are selected fromA-53930A, AE-0047, AGN-190604, AGN-190744, AH-1058, AHR-12742,AHR-16303B, AHR-16462B, AIT-110, AIT-111, AJ-3941, AM-336, amlopidine(including S-(−), R-(+), and racemic), anipamil, AP-1067, aranidipine,atosiban, azelnidipine, barnidipine, Bay-t-7207, Bay-y-5959, Bay-z-4406,BBR-2160, belfosdil, BIII-890-CL, bisaramil, BMS-181102, BMS-188107,BMY-43011, BRL-32872, buflomedil, CD-349, CD-832, CERM-12816, CGP-28932,cilnidipine, clentiazem, clevidipine, CNS-1067, CNS-1237, CNS-2103,CP-060S, CPC-301, CPC-317, CPU-86017, D-2024, darodipine, DHP-218,diltiazem, diperdipine, dopropidil, dotarazine, dronedarone, DTZ-323,E-047/1, efonidipine, EGIS-7229, elgodipine, emopamil, etomoxir, F-0401,fantofarone, fasudil, FCE-24265, FCE-26262, FCE-27335, FCE-27892,FCE-28718, felodipine, FPL-64176, FR-172516, FRG-8701, furnidipine,GS-386, iganidipine, ipenoxazone, isradipine, JTV-591, KP-840, KT-362,L-366682, lacidipine, LAS-0538, LCB-2514, lemildipine, lercanidipine,leualacin, lifarizine, LOE-908, lomerizine, lubeluzole, LY-042826,manidipine, McN-6186, mibefradil, monatepil , MR-14134, N-3601,NCC-1048, nefiracetam, nexopamil, nifedipine, nifedipine, Nifelan,nilvadipine, nimodipine, NNC-09-0026, NPS-568, NS-638, NS-649, NS-696,NS-7, OPC-8490, Org-13061, Org-30029, oxodipine, P-5, palonidipine,PCA-50922, PCA-50938, PCA-50941, PD-029361, PD-157667, PD-158143,PD-176078, pranidipine, QX-314, ranolazine, RHG-2716, RingCap,Ro-11-2933, RS-5773, RU-43945, RWJ-22108, RWJ-22726, RWJ-29009,RWJ-37868, S-12968, S-2150, S-312-d, SANK-71996, SB-201823, SB-206284A,SB-23736, SD-3212, semotiadil, SIB-1281, siratiazem, SKF-45675,SKF-96365, SKT-M-26, SL-34.0829, SL-87.0495, SM-6586, SNX-124, SNX-236,SNX-239, SNX-325, SNX-482, SQ-31727, SQ-33351, SQ-34399, SR-33805,TA-993, tamolarizine, TDN-345, temiverine, terodiline, TH9229, TN-871,U-88999, U-92032, U-92798, UCL-1439, UK-1 656, UK-55444, UK-56593,UK-84149, verapamil, Verelan, vexibinol, VUF-8929, WAY-141520, XB-513,XT-044, Y-22516, YH-334, YM-1615-4, YM-430, Z-6568, zatebradine,ziconotide, and ZM-224832.
 24. A method according to claim 23, whereinthe ligands are selected from the group consisting of verapamil,diltiazem, benziazem clentiazem, nicardipine, nifedipine, nilvadipine,nitredipine, nimodipine, isradipine, lacidipine, amlodipine,nisoldipine, isradipine, mibefrodil, amlodipine, felodipine, nimodipine,bepridil, SQ 32,910 and SQ 32,428.
 25. A method for identifyingmultimeric ligand compounds possessing multibinding properties whichmethod comprises: (a) identifying a ligand or a mixture of ligandswherein each ligand contains at least one reactive functionality; (b)identifying a library of linkers wherein each linker in said librarycomprises at least two functional groups having complementary reactivityto at least one of the reactive functional groups of the ligand; (c)preparing a multimeric ligand compound library by combining at least twostoichiometric equivalents of the ligand or mixture of ligandsidentified in (a) with the library of linkers identified in (b) underconditions wherein the complementary functional groups react to form acovalent linkage between said linker and at least two of said ligands;and (d) assaying the multimeric ligand compounds produced in the libraryprepared in (c) above to identify multimeric ligand compounds possessingmultibinding properties.
 26. A method for identifying multimeric ligandcompounds possessing multibinding properties which method comprises: (a)identifying a library of ligands wherein each ligand contains at leastone reactive functionality; (b) identifying a linker or mixture oflinkers wherein each linker comprises at least two functional groupshaving complementary reactivity to at least one of the reactivefunctional groups of the ligand; (c) preparing a multimeric ligandcompound library by combining at least two stoichiometric equivalents ofthe library of ligands identified in (a) with the linker or mixture oflinkers identified in (b) under conditions wherein the complementaryfunctional groups react to form a covalent linkage between said linkerand at least two of said ligands; and (d) assaying the multimeric ligandcompounds produced in the library prepared in (c) above to identifymultimeric ligand compounds possessing multibinding properties.
 27. Themethod according to claim 25 or 26 wherein the preparation of themultimeric ligand compound library is achieved by either the sequentialor concurrent combination of the two or more stoichiometric equivalentsof the ligands identified in (a) with the linkers identified in (b). 28.The method according to claim 27 wherein the multimeric ligand compoundscomprising the multimeric ligand compound library are dimeric.
 29. Themethod according to claim 28 wherein the dimeric ligand compoundscomprising the dimeric ligand compound library are heterodimeric. 30.The method according to claim 29 wherein the heterodimeric ligandcompound library is prepared by sequential addition of a first andsecond ligand.
 31. The method according to claim 25 or 26 wherein, priorto procedure (d), each member of the multimeric ligand compound libraryis isolated from the library.
 32. The method according to claim 31wherein each member of the library is isolated by preparative liquidchromatography mass spectrometry (LCMS).
 33. The method according toclaim 25 or claim 26 wherein the linker or linkers employed are selectedfrom the group comprising flexible linkers, rigid linkers, hydrophobiclinkers, hydrophilic linkers, linkers of different geometry, acidiclinkers, basic linkers, linkers of different polarization and/orpolarizability and amphiphilic linkers.
 34. The method according toclaim 33 wherein the linkers comprise linkers of different chain lengthand/or having different complementary reactive groups.
 35. The methodaccording to claim 34 wherein the linkers are selected to have differentlinker lengths ranging from about 2 to 100 Å.
 36. The method accordingto claim 25 or 26 wherein the ligand or mixture of ligands is selectedto have reactive functionality at different sites on said ligands. 37.The method according to claim 36 wherein said reactive functionality isselected from the group consisting of carboxylic acids, carboxylic acidhalides, carboxyl esters, amines, halides, pseudohalides, isocyanates,vinyl unsaturation, ketones, aldehydes, thiols, alcohols, anhydrides,boronates, and precursors thereof wherein the reactive functionality onthe ligand is selected to be complementary to at least one of thereactive groups on the linker so that a covalent linkage can be formedbetween the linker and the ligand.
 38. The method according to claim 25or claim 26 wherein the multimeric ligand compound library compriseshomomeric ligand compounds.
 39. The method according to claim 25 orclaim 26 wherein the multimeric ligand compound library comprisesheteromeric ligand compounds.
 40. A library of multimeric ligandcompounds which may possess multivalent properties which library isprepared by the method comprising: (a) identifying a ligand or a mixtureof ligands wherein each ligand contains at least one reactivefunctionality; (b) identifying a library of linkers wherein each linkerin said library comprises at least two functional groups havingcomplementary reactivity to at least one of the reactive functionalgroups of the ligand; and (c) preparing a multimeric ligand compoundlibrary by combining at least two stoichiometric equivalents of theligand or mixture of ligands identified in (a) with the library oflinkers identified in (b) under conditions wherein the complementaryfunctional groups react to form a covalent linkage between said linkerand at least two of said ligands.
 41. A library of multimeric ligandcompounds which may possess multivalent properties which library isprepared by the method comprising: (a) identifying a library of ligandswherein each ligand contains at least one reactive functionality; (b)identifying a linker or mixture of linkers wherein each linker comprisesat least two functional groups having complementary reactivity to atleast one of the reactive functional groups of the ligand; and (c)preparing a multimeric ligand compound library by combining at least twostoichiometric equivalents of the library of ligands identified in (a)with the linker or mixture of linkers identified in (b) under conditionswherein the complementary functional groups react to form a covalentlinkage between said linker and at least two of said ligands.
 42. Thelibrary according to claim 40 or claim 41 wherein the linker or linkersemployed are selected from the group comprising flexible linkers, rigidlinkers, hydrophobic linkers, hydrophilic linkers, linkers of differentgeometry, acidic linkers, basic linkers, linkers of differentpolarization and/or polarizability and amphiphilic linkers.
 43. Thelibrary according to claim 42 wherein the linkers comprise linkers ofdifferent chain length and/or having different complementary reactivegroups.
 44. The library according to claim 43 wherein the linkers areselected to have different linker lengths ranging from about 2 to 100 Å.45. The library according to claim 40 or 41 wherein the ligand ormixture of ligands is selected to have reactive functionality atdifferent sites on said ligands.
 46. The library according to claim 45wherein said reactive functionality is selected from the groupconsisting of carboxylic acids, carboxylic acid halides, carboxylesters, amines, halides, pseudohalides, isocyanates, vinyl unsaturation,ketones, aldehydes, thiols, alcohols, anhydrides, boronates andprecursors thereof wherein the reactive functionality on the ligand isselected to be complementary to at least one of the reactive groups onthe linker so that a covalent linkage can be formed between the linkerand the ligand.
 47. The library according to claim 40 or 41 wherein themultimeric ligand compound library comprises homomeric ligand compounds.48. The library according to claim 40 or 41 wherein the multimericligand compound library comprises heteromeric ligand compounds.
 49. Aniterative method for identifying multimeric ligand compounds possessingmultibinding properties which method comprises: (a) preparing a firstcollection or iteration of multimeric compounds which is prepared bycontacting at least two stoichiometric equivalents of the ligand ormixture of ligands which target a receptor with a linker or mixture oflinkers wherein said ligand or mixture of ligands comprises at least onereactive functionality and said linker or mixture of linkers comprisesat least two functional groups having complementary reactivity to atleast one of the reactive functional groups of the ligand wherein saidcontacting is conducted under conditions wherein the complementaryfunctional groups react to form a covalent linkage between said linkerand at least two of said ligands; (b) assaying said first collection oriteration of multimeric compounds to assess which if any of saidmultimeric compounds possess multibinding properties; (c) repeating theprocess of (a) and (b) above until at least one multimeric compound isfound to possess multibinding properties; (d) evaluating what molecularconstraints imparted multibinding properties to the multimeric compoundor compounds found in the first iteration recited in (a)-(c) above; (e)creating a second collection or iteration of multimeric compounds whichelaborates upon the particular molecular constraints impartingmultibinding properties to the multimeric compound or compounds found insaid first iteration; (f) evaluating what molecular constraints impartedenhanced multibinding properties to the multimeric compound or compoundsfound in the second collection or iteration recited in (e) above; (g)optionally repeating steps (e) and (f) to further elaborate upon saidmolecular constraints.
 50. The method according to claim 49 whereinsteps (e) and (f) are repeated from 2-50 times.
 51. The method accordingto claim 50 wherein steps (e) and (f) are repeated from 5-50 times.